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Wang J, Qian X, Yin Z, Tong D, Wang C, Hu Z, Cai W, Tang J. Characterization of hepatocarcinoma cells: lymphatic metastasis potential through enhanced production, expression, and secretion along the VEGF-C/D-VEGFR-3/NRP-2 axis. Discov Oncol 2025; 16:191. [PMID: 39960569 PMCID: PMC11832830 DOI: 10.1007/s12672-025-01901-z] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 08/05/2024] [Accepted: 02/03/2025] [Indexed: 02/20/2025] Open
Abstract
OBJECTIVES In vitro and in vivo analyses were conducted to examine the correlations between the production, expression, and secretion of the VEGF-C/D-VEGFR-3/NRP-2 axis and various levels of lymphatic metastatic potential in murine hepatocellular carcinoma cells. METHODS Quantitative real-time PCR, western blotting, cytoimmunofluorescence, immunohistochemistry, and enzyme-linked immunosorbent assay were employed to assess the expression and secretion of the VEGF-C/D-VEGFR-3/NRP-2 axis at the gene, protein, cytological, and histological levels. RESULTS In both in vitro and in vivo experiments, the ligands VEGF-C/D and receptors VEGFR-3/NRP-2 were primarily localized in the cytoplasm or cell membrane of hepatocarcinoma F/P cells with high/low lymphatic metastatic potentials, and in normal hepatocytes, respectively. The production and expression levels of the VEGF-C/D-VEGFR-3/NRP-2 axis in F/P cells were significantly higher than those in normal liver cells. Additionally, the production and expression of this axis were higher in F cells compared to P cells (P < 0.01). Notably, within both F/P cells, the production and expression levels of VEGF-C and VEGFR-3 were lower than those of VEGF-D and NRP-2 (P < 0.05). However, the secretion of VEGF-C exceeded that of VEGF-D in the supernatant of cultured cells and in the serum of tumor-bearing mice (P < 0.01). Interestingly, the ratio of VEGF-C/D in F cells was significantly higher than that in P cells and normal hepatocytes. Moreover, the ratio of VEGF-C/D was notably elevated in the supernatant of tumor cells and serum from tumor-bearing mice, as determined by ELISA, compared to that in tumor cells and tissues assessed using other technologies (P < 0.01). CONCLUSION The significant co-production, co-expression, and co-secretion of the VEGF-C/D-VEGFR-3/NRP-2 axis are distinctive features of hepatocarcinoma cells, particularly those with a heightened potential for lymphatic metastasis.
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Affiliation(s)
- Jingwen Wang
- Department of Pathology, Tianjin First Central Hospital, School of Medicine, Nankai University, No.2 Baoshan Road, Xiqing District, Tianjin, 300192, China
| | - Xuejiao Qian
- Department of Respiratory Medicine, Tianjin First Central Hospital, School of Medicine, Nankai University, Tianjin, 300192, China
| | - Zhiqi Yin
- Department of Pathology, Tianjin First Central Hospital, School of Medicine, Nankai University, No.2 Baoshan Road, Xiqing District, Tianjin, 300192, China
| | - Dan Tong
- Department of Pathology, Tianjin First Central Hospital, School of Medicine, Nankai University, No.2 Baoshan Road, Xiqing District, Tianjin, 300192, China
| | - Caihong Wang
- Department of Pathology, Tianjin First Central Hospital, School of Medicine, Nankai University, No.2 Baoshan Road, Xiqing District, Tianjin, 300192, China
| | - Zhandong Hu
- Department of Pathology, Tianjin First Central Hospital, School of Medicine, Nankai University, No.2 Baoshan Road, Xiqing District, Tianjin, 300192, China
| | - Wenjuan Cai
- Department of Pathology, Tianjin First Central Hospital, School of Medicine, Nankai University, No.2 Baoshan Road, Xiqing District, Tianjin, 300192, China.
| | - Jianwu Tang
- Department of Pathology, Key Laboratory for Tumor Metastasis and Intervention of Liaoning Province, Dalian Medical University, 9 West, Lvshun Southern Road, Dalian Liaoning, 116044, China.
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Vimalraj S, Hariprabu KNG, Rahaman M, Govindasami P, Perumal K, Sekaran S, Ganapathy D. Vascular endothelial growth factor-C and its receptor-3 signaling in tumorigenesis. 3 Biotech 2023; 13:326. [PMID: 37663750 PMCID: PMC10474002 DOI: 10.1007/s13205-023-03719-4] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/23/2022] [Accepted: 07/13/2023] [Indexed: 09/05/2023] Open
Abstract
The cancer-promoting ligand vascular endothelial growth factor-C (VEGF-C) activates VEGF receptor-3 (VEGFR-3). The VEGF-C/VEGFR-3 axis is expressed by a range of human tumor cells in addition to lymphatic endothelial cells. Activating the VEGF-C/VEGFR-3 signaling enhances metastasis by promoting lymphangiogenesis and angiogenesis inside and around tumors. Stimulation of VEGF-C/VEGFR-3 signaling promotes tumor metastasis in tumors, such as ovarian, renal, pancreatic, prostate, lung, skin, gastric, colorectal, cervical, leukemia, mesothelioma, Kaposi sarcoma, and endometrial carcinoma. We discuss and update the role of VEGF-C/VEGFR-3 signaling in tumor development and the research is still needed to completely comprehend this multifunctional receptor.
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Affiliation(s)
- Selvaraj Vimalraj
- Department of Applied Mechanics and Biomedical Engineering, Indian Institute of Technology, Madras, Chennai, India
| | | | - Mostafizur Rahaman
- Department of Chemistry, College of Science, King Saud University, P. O. Box 2455, Riyadh, 11451 Saudi Arabia
| | - Periyasami Govindasami
- Department of Chemistry, College of Science, King Saud University, P. O. Box 2455, Riyadh, 11451 Saudi Arabia
| | - Karthikeyan Perumal
- Department of Chemistry and Biochemistry, The Ohio State University, 151 W. Woodruff Ave, Columbus, OH 43210 USA
| | - Saravanan Sekaran
- Department of Prosthodontics, Saveetha Dental College and Hospital, Saveetha Institute of Medical and Technical Sciences (SIMATS), Saveetha University, Chennai, Tamil Nadu 600 077 India
| | - Dhanraj Ganapathy
- Department of Prosthodontics, Saveetha Dental College and Hospital, Saveetha Institute of Medical and Technical Sciences (SIMATS), Saveetha University, Chennai, Tamil Nadu 600 077 India
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Ai H. GSEA-SDBE: A gene selection method for breast cancer classification based on GSEA and analyzing differences in performance metrics. PLoS One 2022; 17:e0263171. [PMID: 35472078 PMCID: PMC9041804 DOI: 10.1371/journal.pone.0263171] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/23/2021] [Accepted: 01/13/2022] [Indexed: 12/20/2022] Open
Abstract
MOTIVATION Selecting the most relevant genes for sample classification is a common process in gene expression studies. Moreover, determining the smallest set of relevant genes that can achieve the required classification performance is particularly important in diagnosing cancer and improving treatment. RESULTS In this study, I propose a novel method to eliminate irrelevant and redundant genes, and thus determine the smallest set of relevant genes for breast cancer diagnosis. The method is based on random forest models, gene set enrichment analysis (GSEA), and my developed Sort Difference Backward Elimination (SDBE) algorithm; hence, the method is named GSEA-SDBE. Using this method, genes are filtered according to their importance following random forest training and GSEA is used to select genes by core enrichment of Kyoto Encyclopedia of Genes and Genomes pathways that are strongly related to breast cancer. Subsequently, the SDBE algorithm is applied to eliminate redundant genes and identify the most relevant genes for breast cancer diagnosis. In the SDBE algorithm, the differences in the Matthews correlation coefficients (MCCs) of performing random forest models are computed before and after the deletion of each gene to indicate the degree of redundancy of the corresponding deleted gene on the remaining genes during backward elimination. Next, the obtained MCC difference list is divided into two parts from a set position and each part is respectively sorted. By continuously iterating and changing the set position, the most relevant genes are stably assembled on the left side of the gene list, facilitating their identification, and the redundant genes are gathered on the right side of the gene list for easy elimination. A cross-comparison of the SDBE algorithm was performed by respectively computing differences between MCCs and ROC_AUC_score and then respectively using 10-fold classification models, e.g., random forest (RF), support vector machine (SVM), k-nearest neighbor (KNN), extreme gradient boosting (XGBoost), and extremely randomized trees (ExtraTrees). Finally, the classification performance of the proposed method was compared with that of three advanced algorithms for five cancer datasets. Results showed that analyzing MCC differences and using random forest models was the optimal solution for the SDBE algorithm. Accordingly, three consistently relevant genes (i.e., VEGFD, TSLP, and PKMYT1) were selected for the diagnosis of breast cancer. The performance metrics (MCC and ROC_AUC_score, respectively) of the random forest models based on 10-fold verification reached 95.28% and 98.75%. In addition, survival analysis showed that VEGFD and TSLP could be used to predict the prognosis of patients with breast cancer. Moreover, the proposed method significantly outperformed the other methods tested as it allowed selecting a smaller number of genes while maintaining the required classification accuracy.
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Affiliation(s)
- Hu Ai
- Department of Criminal Technology, Guizhou Police College, Guiyang, Guizhou, China
- * E-mail:
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Hegde M, Daimary UD, Kumar A, Chinnathambi A, Alharbi SA, Shakibaei M, Kunnumakkara AB. STAT3/HIF1A and EMT specific transcription factors regulated genes: Novel predictors of breast cancer metastasis. Gene X 2022; 818:146245. [PMID: 35074419 DOI: 10.1016/j.gene.2022.146245] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/03/2021] [Accepted: 01/18/2022] [Indexed: 12/26/2022] Open
Abstract
Metastasis, the fatal hallmark of breast cancer (BC), is a serious hurdle for therapy. Current prognostic approaches are not sufficient to predict the metastasis risk for BC patients. Therefore, in the present study, we analyzed gene expression data from GSE139038 and TCGA database to develop predictive markers for BC metastasis. Initially, the data from GSE139038 which contained 65 samples consisting of 41 breast tumor tissues, 18 paired morphologically normal tissues and 6 from non-malignant breast tissues were analyzed for differentially expressed genes (DEGs). DEGs were obtained from three different comparisons: paired morphologically normal (MN) versus tumor samples (C), apparently normal (AN) versus tumor samples (C), and paired morphologically normal (MN) versus apparently normal samples (AN). Multiple bioinformatic methods were employed to evaluate metastasis, EMT and triple negative breast cancer (TNBC) specific genes. Further, regulation of gene expression, clinicopathological factors and DNA methylation patterns of DEGs in BC were validated with TCGA datasets. Our bioinformatic analysis showed that 40 genes were upregulated and 294 were found to be downregulated between AN vs C; 124 were upregulated and 760 genes were downregulated between MN vs C; 4 were upregulated and 13 were downregulated between MN vs AN. Analysis using TCGA dataset revealed 18 genes were significantly altered in nodal positive BC patients compared to nodal negative BC patients. Our study showed novel candidate genes as predictive markers for BC metastasis which can also be used for therapeutic targets for BC treatment.
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Affiliation(s)
- Mangala Hegde
- Cancer Biology Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology-Guwahati, Guwahati 781 039, Assam, India; DBT-AIST International Center for Translational and Environmental Research, Indian Institute of Technology-Guwahati, Guwahati 781 039, Assam, India
| | - Uzini Devi Daimary
- Cancer Biology Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology-Guwahati, Guwahati 781 039, Assam, India; DBT-AIST International Center for Translational and Environmental Research, Indian Institute of Technology-Guwahati, Guwahati 781 039, Assam, India
| | - Aviral Kumar
- Cancer Biology Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology-Guwahati, Guwahati 781 039, Assam, India; DBT-AIST International Center for Translational and Environmental Research, Indian Institute of Technology-Guwahati, Guwahati 781 039, Assam, India
| | - Arunachalam Chinnathambi
- Department of Botany and Microbiology, College of Science, King Saud University, PO Box 2455, Riyadh 11451, Saudi Arabia
| | - Sulaiman Ali Alharbi
- Department of Botany and Microbiology, College of Science, King Saud University, PO Box 2455, Riyadh 11451, Saudi Arabia
| | - Mehdi Shakibaei
- Musculoskeletal Research Group and Tumor Biology, Chair of Vegetative Anatomy, Faculty of Medicine, Institute of Anatomy, Ludwig-Maximilian-University Munich, Munich, Germany
| | - Ajaikumar B Kunnumakkara
- Cancer Biology Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology-Guwahati, Guwahati 781 039, Assam, India; DBT-AIST International Center for Translational and Environmental Research, Indian Institute of Technology-Guwahati, Guwahati 781 039, Assam, India.
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Genetic Variations of Kinase Inserts Domain Receptor (KDR) Gene Are Associated with the Risk of Astrocytomas. Mol Neurobiol 2015; 53:2541-9. [PMID: 26081139 DOI: 10.1007/s12035-015-9239-6] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/02/2014] [Accepted: 05/26/2015] [Indexed: 02/08/2023]
Abstract
Astrocytomas is one of the most common central nervous system (CNS) tumors with high mortality rate. Kinase insert domain receptor (KDR) is involved in the regulation of tumor angiogenesis, migration, and vascular permeability. The aim of the study was to explore the relationship between KDR polymorphisms and risk of astrocytomas. Blood samples were collected from 157 astrocytomas patients and 160 healthy controls. Three tag-SNPs (rs2071559C/T, rs2305948T/C, and rs1870377A/T) were identified from the International HapMap Project Databases and genotyped using the method of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). We evaluated the astrocytomas risk caused by individual SNPs and haplotype using odds ratios (ORs) and their 95 % confidence intervals (CIs). In the overall individual SNP analysis, the C allele of rs2071559 was correlated with an increased risk of astrocytomas. However, individuals with mutant allele A and genotype TA + AA of rs1870377 showed a protective effect against astrocytomas. Subgroup analysis based on WHO tumor grade revealed that the C allele of rs2071559 had more influence with the risk of astrocytomas in the grade III-IV (OR = 1.91) subgroup than the grade I-II (OR = 1.47) group. Genotype TT of rs2305948 was found to be significantly associated with susceptibility of astrocytomas only in the grade III-IV subgroup. The protective effect of rs1870377 did not reveal significant difference between the grade III-IV and grade I-II subgroups. Meanwhile, stratified analysis demonstrated that mutation of rs2071559 and rs2305948 could elevate the risk of astrocytomas more significantly in the subgroup of smokers than the nonsmokers. Interestingly, the protective effect of rs1870377 was more obvious in the nonsmokers than the smokers. Additionally, haplotype-specific analysis showed that haplotype CCT and CTT were related with an increased risk of astrocytomas. We found that individual with variants of rs2071559*C and rs2305948*T might significantly elevate the risk of astrocytomas, while mutants of rs1870377*A was associated with the decreased risk of astrocytomas. Further studies about ethnically diverse populations with larger sample size should be performed to confirm the correlation between KDR gene polymorphisms and risk of astrocytomas.
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Wang CA, Tsai SJ. The non-canonical role of vascular endothelial growth factor-C axis in cancer progression. Exp Biol Med (Maywood) 2015; 240:718-24. [PMID: 25888649 DOI: 10.1177/1535370215583802] [Citation(s) in RCA: 26] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022] Open
Abstract
It has been shown in many clinical studies that the level of vascular endothelial growth factor-C (VEGF-C) positively correlates with lymph node metastasis. Nevertheless, beyond the canonical role of VEGF-C in stimulating lymphangiogenesis and thus promoting lymph node/distant metastasis, emerging evidence indicates that expression of VEGF-C contributes to various aspects of carcinogenicity via autocrine regulation. The newly identified functions of VEGF-C include but are not limited to proliferation, migration, invasion, and chemo-resistance. Besides tumor cell autocrine regulation, VEGF-C can also modulate the immune system such that tumor cells more easily escape immune surveillance. Therefore, understanding the functional roles and regulatory mechanisms related to the VEGF-C axis may lead to alternative strategies for cancer treatment. This mini-review will focus on summarizing recent discoveries regarding the unconventional functions of VEGF-C in cancer progression.
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Affiliation(s)
- Chu-An Wang
- Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan 70101, Taiwan
| | - Shaw-Jenq Tsai
- Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan 70101, Taiwan Department of Physiology, College of Medicine, National Cheng Kung University, Tainan 70101, Taiwan
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Gong S, Seng Z, Wang W, Lv J, Dong Q, Yan B, Peng L, He X. Bosentan protects the spinal cord from ischemia reperfusion injury in rats through vascular endothelial growth factor receptors. Spinal Cord 2014; 53:19-23. [PMID: 25179655 DOI: 10.1038/sc.2014.147] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/12/2014] [Revised: 07/09/2014] [Accepted: 07/28/2014] [Indexed: 11/09/2022]
Abstract
STUDY DESIGN Experimental study. OBJECTIVES To investigate whether Bosentan, an endothelin-A/-B dual receptor antagonist, could protect neurons after spinal cord ischemia reperfusion (SCIR) injury in rats and its underlying signaling pathway. SETTING Department of Neurosurgery, the Second Affiliated Hospital, Xi'an Jiaotong University School of Medicine, Xi'an, Shaanxi Province, China. METHODS Sprague-Dawley rats were randomly divided into two groups, saline group (IRS, n=48) and Bosentan group (IRB, 5 mg kg(-1), n=48). After ischemia for 1 h with occlusion of the infrarenal aorta, spinal cord were reperfused for 6h, 12h, 24h, 3d, 5d, and 7d separately. Enzyme-linked immunosorbent assay was used to detect vascular endothelial growth factor (VEGF) in serum. Immunohistochemistry was performed to detect protein expression of VEGF, VEGF receptor 1 (FLT-1) and VEGF receptor 2 (FLK-1). Gene expressions of VEGF and its receptors were evaluated using the quantitative reverse transcription polymerase chain reaction. RESULTS Compared with the IRS group, gene and protein expressions of VEGF, FLT-1 and FLK-1 were significantly increased (P<0.05), so was the concentration of VEGF in plasma (P<0.05). FLK-1 was expressed on spinal cord neurons.
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Affiliation(s)
- S Gong
- Department of Neurosurgery, The Second Affiliated Hospital, Xi'an Jiaotong University School of Medicine, Shaanxi Province, China
| | - Z Seng
- Department of Neurosurgery, The Second Affiliated Hospital, Xi'an Jiaotong University School of Medicine, Shaanxi Province, China
| | - W Wang
- Department of Spine Surgery, Xi'an Red Cross Society Hospital, Xi'an Jiaotong University, Shaanxi Province, China
| | - J Lv
- Department of Neurosurgery, The Second Affiliated Hospital, Xi'an Jiaotong University School of Medicine, Shaanxi Province, China
| | - Q Dong
- Department of Neurosurgery, The Second Affiliated Hospital, Xi'an Jiaotong University School of Medicine, Shaanxi Province, China
| | - B Yan
- Department of Emergency Medicine, The Second Affiliated Hospital, Xi'an Jiaotong University School of Medicine, Shaanxi Province, China
| | - L Peng
- Department of Cardiology, The Second Affiliated Hospital, Xi'an Jiaotong University School of Medicine, Shaanxi Province, China
| | - X He
- Department of Orthopaedics, The Second Affiliated Hospital, Xi'an Jiaotong University School of Medicine, Shaanxi Province, China
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de Sousa EA, Lourenço SV, de Moraes FPP, Vartanian JG, Gonçalves-Filho J, Kowalski LP, Soares FA, Coutinho-Camillo CM. Head and neck squamous cell carcinoma lymphatic spread and survival: Relevance of vascular endothelial growth factor family for tumor evaluation. Head Neck 2014; 37:1410-6. [PMID: 24824527 DOI: 10.1002/hed.23765] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/11/2013] [Revised: 03/14/2014] [Accepted: 05/10/2014] [Indexed: 01/06/2023] Open
Abstract
BACKGROUND Head and neck squamous cell carcinoma (HNSCC) is primarily a locoregional disease in which the cervical lymph nodes are the chief site of metastasis. The purpose of this study was to examine the relationship between lymphangiogenesis and clinicopathological aspects of HNSCC and its metastasis. METHODS Fifty-two patients with HNSCC and metastatic lymph nodes from 21 of these subjects were analyzed by immunohistochemistry. RESULTS The HNSCC samples were predominantly negative for vascular endothelial growth factor (VEGF)-C, VEGF-D, and vascular endothelial growth factor receptor (VEGFR)3. There was an association between the density of lymph vessels (measured by D2-40 staining) in the lymph nodes and advanced-stage tumors. There was no link between the expression of these proteins and survival rates. CONCLUSION Although lymphatic spread is a significant event in the progression of HNSCC, the expression of VEGF-C, VEGF-D, and VEGFR3 does not correlate with clinicopathological characteristics, suggesting that other signaling pathways mediate lymphangiogenesis in HNSCC.
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Affiliation(s)
| | | | | | - José Guilherme Vartanian
- Department of Head and Neck Surgery and Otorhinolaryngology, AC Camargo Cancer Center, São Paulo, Brazil
| | - João Gonçalves-Filho
- Department of Head and Neck Surgery and Otorhinolaryngology, AC Camargo Cancer Center, São Paulo, Brazil
| | - Luiz Paulo Kowalski
- Department of Head and Neck Surgery and Otorhinolaryngology, AC Camargo Cancer Center, São Paulo, Brazil
| | - Fernando Augusto Soares
- Department of Pathology, AC Camargo Cancer Center, São Paulo, Brazil.,Department of General Pathology, Dental School, University of São Paulo, Brazil
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Stacker SA, Williams SP, Karnezis T, Shayan R, Fox SB, Achen MG. Lymphangiogenesis and lymphatic vessel remodelling in cancer. Nat Rev Cancer 2014; 14:159-72. [PMID: 24561443 DOI: 10.1038/nrc3677] [Citation(s) in RCA: 579] [Impact Index Per Article: 52.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
The generation of new lymphatic vessels through lymphangiogenesis and the remodelling of existing lymphatics are thought to be important steps in cancer metastasis. The past decade has been exciting in terms of research into the molecular and cellular biology of lymphatic vessels in cancer, and it has been shown that the molecular control of tumour lymphangiogenesis has similarities to that of tumour angiogenesis. Nevertheless, there are significant mechanistic differences between these biological processes. We are now developing a greater understanding of the specific roles of distinct lymphatic vessel subtypes in cancer, and this provides opportunities to improve diagnostic and therapeutic approaches that aim to restrict the progression of cancer.
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Affiliation(s)
- Steven A Stacker
- 1] Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia. [2] Sir Peter MacCallum Department of Oncology, University of Melbourne, Victoria 3010, Australia. [3] Department of Surgery, Royal Melbourne Hospital, University of Melbourne, Parkville, Victoria 3050, Australia
| | - Steven P Williams
- Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia
| | - Tara Karnezis
- 1] Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia. [2] Sir Peter MacCallum Department of Oncology, University of Melbourne, Victoria 3010, Australia
| | - Ramin Shayan
- 1] Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia. [2] Department of Surgery, Royal Melbourne Hospital, University of Melbourne, Parkville, Victoria 3050, Australia. [3] Department of Surgery, St. Vincent's Hospital, University of Melbourne, Fitzroy, Victoria 3065, Australia. [4] O'Brien Institute, Australian Catholic University, Fitzroy, Victoria 3065, Australia
| | - Stephen B Fox
- 1] Sir Peter MacCallum Department of Oncology, University of Melbourne, Victoria 3010, Australia. [2] Department of Pathology, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia
| | - Marc G Achen
- 1] Tumour Angiogenesis Program, Peter MacCallum Cancer Centre, East Melbourne, Victoria 3002, Australia. [2] Sir Peter MacCallum Department of Oncology, University of Melbourne, Victoria 3010, Australia. [3] Department of Surgery, Royal Melbourne Hospital, University of Melbourne, Parkville, Victoria 3050, Australia
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Wang Z, Wu XL, Wang X, Tian HX, Chen ZH, Li YQ. The biophysical property of A549 cells transferred by VEGF-D. SCANNING 2014; 36:202-208. [PMID: 23526563 DOI: 10.1002/sca.21087] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/16/2013] [Revised: 02/02/2013] [Accepted: 02/11/2013] [Indexed: 06/02/2023]
Abstract
Vascular endothelial growth factor-D (VEGF-D) together with VEGF-C is considered to be associated with lymphangiogenesis and angiogenesis and involve in tumorization. This study aims to investigate the influence of exogenous VEGF-D gene on the biophysical property of cell surface of lung adenocarcinoma cell line. A panel of lung adenocarcinoma cell lines were examined the expression of VEGF-D and VEGF-C by real-time PCR. The VEGF-D recombinant plasmid containing enhanced green fluorescence protein (EGFP) was constructed and transfected to the cell line with no expression of VEGF-D and confirmed by real-time PCR and Western blot analysis. Topographic images of cells were obtained by using atomic force microscope (AFM) in contact mode. Unlike VEGF-C, VEGF-D was found to have a very low expression or undetectable expression in lung adenocarcinoma cell lines. The VEGF-D recombinant plasmid had been constructed successfully and was transferred into the human lung adenocarcinoma cell line A549 cells which had no endogenous expression of VEGF-D, and exogenous VEGF-D could be detected in mRNA and protein expression levels in the gene modified cells, while the VEGF-C gene expression had no change after VEGF-D transfection. After transfection, the irregular microspikes or nano clusters could observe on the surface of A549 cells, and VEGF-D transfected A549 cells became more rigid. The exogenous VEGF-D gene might cause the remarkable biophysical architectural changes in the A549 cells, which might as a novel biomarker for evaluation of its biological function.
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Affiliation(s)
- Zhen Wang
- Guangdong Lung Cancer Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, PR China
| | - Xiu-Li Wu
- Institute of Hematology, Medical College, Jinan University, Guangzhou, PR China
| | - Xu Wang
- Key Laboratory for Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou, PR China
| | - Hong-Xia Tian
- Guangdong Lung Cancer Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, PR China
| | - Zhi-Hong Chen
- Guangdong Lung Cancer Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, PR China
| | - Yang-Qiu Li
- Institute of Hematology, Medical College, Jinan University, Guangzhou, PR China
- Key Laboratory for Regenerative Medicine of Ministry of Education, Jinan University, Guangzhou, PR China
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He L, He J, Zhao X. Expression of VEGF-D in epithelial ovarian cancer and its relationship to lymphatic metastasis. Asia Pac J Clin Oncol 2013; 12:e161-6. [PMID: 23915006 DOI: 10.1111/ajco.12086] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
Abstract
AIM To investigate the contribution of vascular endothelial growth factor (VEGF)-D to tumor progression, tumor lymphangiogenesis and lymphatic metastasis in epithelial ovarian cancer. METHODS The expression profiles of VEGF-D in 18 benign, 14 borderline and 87 malignant epithelial ovarian cancers were examined using immunohistochemical (IHC) staining. Lymphatic vessels were identified using IHC staining on lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), which is a lymph-specific receptor for hyaluronan in identifying lymphatic vessels. The potential correlation among VEGF-D, lymphatic vessel density (LVD) and clinico-pathological factors of the epithelial ovarian cancer was also analyzed. RESULTS Positive IHC staining of VEGF-D was observed in 17% of benign, 21% of borderline and 80% of malignant epithelial ovarian tumors specimens. In the epithelial ovarian cancer specimens, the LVD was 3.41 ± 2.37 in the VEGF-D negative (17 patients), 5.42 ± 3.49 in the weak (26 patients), 7.22 ± 2.36 in the moderate (27 patients) and 7.35 ± 4.06 in the strong (17 patients) groups, respectively. Additionally, the expression of VEGF-D was positively correlated with LVD (r = 0.415, P < 0.001). The expression level of VEGF-D was significantly higher in lymph node-positive epithelial ovarian cancer than in lymph node-negative patients (P = 0.009, P < 0.05). The expression of VEGF-D was significantly correlated with lymph node metastasis, International Federation of Gynecology and Obstetrics stage and tumor histological differentiation, but not with the patients' age or histology type. CONCLUSION VEGF-D may play an important role in the process of lymphatic metastasis of epithelial ovarian cancer.
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Affiliation(s)
- Lixia He
- Department of Obstetrics and Gynecology, West China Second University Hospital of Sichuan University, Chengdu, China
| | - Junyong He
- Physical Examination Center, West China Hospital, Sichuan University, Chengdu, China
| | - Xia Zhao
- Department of Obstetrics and Gynecology, West China Second University Hospital of Sichuan University, Chengdu, China
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Semenza GL. Cancer-stromal cell interactions mediated by hypoxia-inducible factors promote angiogenesis, lymphangiogenesis, and metastasis. Oncogene 2012; 32:4057-63. [PMID: 23222717 DOI: 10.1038/onc.2012.578] [Citation(s) in RCA: 163] [Impact Index Per Article: 12.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/10/2012] [Accepted: 10/09/2012] [Indexed: 12/13/2022]
Abstract
Interactions between cancer cells and stromal cells, including blood vessel endothelial cells (BECs), lymphatic vessel endothelial cells (LECs), bone marrow-derived angiogenic cells (BMDACs) and other bone marrow-derived cells (BMDCs) play important roles in cancer progression. Intratumoral hypoxia, which affects both cancer and stromal cells, is associated with a significantly increased risk of metastasis and mortality in many human cancers. Recent studies have begun to delineate the molecular mechanisms underlying the effect of intratumoral hypoxia on cancer progression. Reduced O2 availability induces the activity of hypoxia-inducible factors (HIFs), which activate the transcription of target genes encoding proteins that play important roles in many critical aspects of cancer biology. Included among these are secreted factors, including angiopoietin 2, angiopoietin-like 4, placental growth factor, platelet-derived growth factor B, stem cell factor (kit ligand), stromal-derived factor 1, and vascular endothelial growth factor. These factors are produced by hypoxic cancer cells and directly mediate functional interactions with BECs, LECs, BMDACs and other BMDCs that promote angiogenesis, lymphangiogenesis, and metastasis. In addition, lysyl oxidase (LOX) and LOX-like proteins, which are secreted by hypoxic breast cancer cells, remodel extracellular matrix in the lungs, which leads to BMDC recruitment and metastatic niche formation.
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Affiliation(s)
- G L Semenza
- Vascular Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
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Wang J, Guo Y, Wang B, Bi J, Li K, Liang X, Chu H, Jiang H. Lymphatic microvessel density and vascular endothelial growth factor-C and -D as prognostic factors in breast cancer: a systematic review and meta-analysis of the literature. Mol Biol Rep 2012; 39:11153-65. [PMID: 23054001 DOI: 10.1007/s11033-012-2024-y] [Citation(s) in RCA: 38] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/17/2012] [Accepted: 10/02/2012] [Indexed: 12/31/2022]
Abstract
The use of lymphatic microvessel density (LVD) and pro-lymphangiogenic mediators as prognostic factors for survival in breast cancer remains controversial. We searched the electronic databases PubMed and EMBASE without language restrictions for relevant literature to aggregate the survival results. To be eligible, every study had to include the assessment of the LVD or the expression of vascular endothelial growth factor (VEGF)-C or -D in patients with breast cancer and provide a survival comparison, including disease-free survival (DFS) or overall survival (OS), according to the LVD, VEGF-C or VEGF-D status. Across all studies, 56.64 % of patients were considered to have a VEGF-C-positive tumor, and 65.54 % of patients had VEGF-D-positive tumors. High LVD had an unfavorable impact on DFS, with a pooled hazard ratio (HR) of 2.222 (95 % CI 1.579-3.126) and an OS with a HR of 2.493 (95 % CI 1.183-5.25). According to the different lymphatic makers, the subgroup HR in the D2-40 studies was 2.431 (95 % CI 1.622-3.644) for DFS and 4.085 (95 % CI 1.896-8.799) for OS. VEGF-C overexpression, as assessed by immunochemistry, was a prognostic factor for decreased DFS (HR 2.164; 95 % CI 1.256-3.729) and for decreased OS (HR 2.613; 95 % CI 1.637-4.170). VEGF-D overexpression was a significant although weak prognostic factor for DFS only when assessed by immunochemistry, with a HR of 2.108 (95 % CI 1.014-4.384). Our meta-analysis demonstrated that LVD, VEGF-C and VEGF-D could predict poor prognosis in patients with breast cancer. However, standardization of the assessment of LVD and for the expression of lymphangiogenesis factors is needed.
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Affiliation(s)
- Jun Wang
- Department of Oncology, General Hospital, Jinan Command of People's Liberation Army, Shifan Street 25, Tianqiao District, Jinan, 250031, China.
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Hypoxia-inducible factor 1-dependent expression of platelet-derived growth factor B promotes lymphatic metastasis of hypoxic breast cancer cells. Proc Natl Acad Sci U S A 2012; 109:E2707-16. [PMID: 23012449 DOI: 10.1073/pnas.1214019109] [Citation(s) in RCA: 167] [Impact Index Per Article: 12.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/18/2022] Open
Abstract
Lymphatic dissemination from the primary tumor is a major mechanism by which breast cancer cells access the systemic circulation, resulting in distant metastasis and mortality. Numerous studies link activation of hypoxia-inducible factor 1 (HIF-1) with tumor angiogenesis, metastasis, and patient mortality. However, the role of HIF-1 in lymphatic dissemination is poorly understood. In this study, we show that HIF-1 promotes lymphatic metastasis of breast cancer by direct transactivation of the gene encoding platelet-derived growth factor B (PDGF-B), which has proliferative and chemotactic effects on lymphatic endothelial cells. Lymphangiogenesis and lymphatic metastasis in mice bearing human breast cancer orthografts were blocked by administration of the HIF-1 inhibitor digoxin or the tyrosine kinase inhibitor imatinib. Immunohistochemical analysis of human breast cancer biopsies demonstrated colocalization of HIF-1α and PDGF-B, which were correlated with lymphatic vessel area and histological grade. Taken together, these data provide experimental support for breast cancer clinical trials targeting HIF-1 and PDGF-B.
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Abstract
Historically, lymphatic vessels were considered passive participants in tumor metastasis by simply providing channels for tumor cells to transit to draining lymph nodes. The discovery of several key lymphatic-specific molecular markers and an increased availability of in vitro and in vivo experimental systems to study lymphatic biology have however highlighted a much more complex, active role for the lymphatic vasculature in metastatic tumor spread. This review will briefly describe the lymphatic system and lymphangiogenesis and then focus on the role of the lymphatic system in cancer metastasis. The progression of our understanding from the lymphatic system as a somewhat passive conduit for metastasis to an active participant in metastatic tumor dissemination, regulated by a complex array of lymphangiogenic factors, chemokines, and immune cell subsets, will be described.
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Affiliation(s)
- Ailsa Christiansen
- Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology, ETH Zurich, Zurich, Switzerland
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16
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Cai X, Ma S, Gu M, Zu C, Qu W, Zheng X. Survivin regulates the expression of VEGF-C in lymphatic metastasis of breast cancer. Diagn Pathol 2012; 7:52. [PMID: 22607367 PMCID: PMC3487795 DOI: 10.1186/1746-1596-7-52] [Citation(s) in RCA: 20] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/11/2012] [Accepted: 05/08/2012] [Indexed: 01/11/2023] Open
Abstract
Background As a known regulator of apoptosis, survivin has positive relationship with lymphatic metastasis in breast cancer. This study aims to detect the difference in expression between survivin and vascular endothelial growth factor-C (VEGF-C) in treated breast cancer cells and tissues, and to analyze the correlation among survivin, VEGF-C and lymphatic metastasis. Methods Plasmid with survivin and VEGF-C shRNA and lentivirus with survivin gene were constructed and transfected into breast cancer cell ZR-75-30. Then the expressions of the two genes were examined using western blot analysis and real-time PCR. The change of invasiveness of breast cancer cells was assessed using matrigel invasion assay. Using immunohistochemistry, the expression of survivin and VEGF-C were analyzed in 108 clinical breast cancer cases with breast cancer tissue and lymph node. Results Survivin regulated the expression of VEGF-C at both protein and mRNA levels in breast cancer cells. Immunohistochemical analysis showed that the level of VEGF-C expression was significantly related with that of survivin in breast cancer tissues (p<0.05). VEGF-C was found to participate in the process of breast cancer cells invasion mediated by survivin. The co-expression of the two and the single expression of any one took significant difference in positive lymph node (p<0.05). Conclusions Survivin takes an important part in regulating the expression of VEGF-C. VEGF-C could influence the invasive ability mediated by survivin. The co-expression of survivin and VEGF-C is more statistically significant to assess lymphatic metastasis in breast cancer. Virtual slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/9193530897100952
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Affiliation(s)
- Xiaopeng Cai
- Department of Breast Surgery, First Affiliated Hospital, China Medical University, No, 155 North Nanjing Street, Shenyang, Liaoning Province, 110001, China
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Zhu P, Zhang J, Chen Q, Wang J, Wang Y. Expression of vascular endothelial growth factor-C in gastric carcinoma and the effect of its antisense gene transfection on the proliferation of human gastric cancer cell line SGC-7901. Am J Surg 2012; 204:78-83. [PMID: 22227171 DOI: 10.1016/j.amjsurg.2011.06.056] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/27/2010] [Revised: 06/28/2011] [Accepted: 06/28/2011] [Indexed: 01/22/2023]
Abstract
PURPOSE The aim of this study was to investigate the relationship between the expression of vascular endothelial growth factor-C (VEGF-C) in gastric carcinoma and tumor lymphangiogenesis and to determine the effect of antisense-VEGF-C gene transfection on proliferation. METHODS Adjacent cancer tissues were collected from 72 gastric carcinoma cases and compared with 10 nongastric carcinoma tissues to detect the expression of VEGF-C and its messenger RNA (mRNA) and calculate the density of neonatal lymphatic microvessels. The in vitro-cultured gastric cancer cell line SGC-7901 was transfected with recombinant plasmid pCI-neo-anti VEGF-C. The expression in the transfected cells and the proliferation were determined. RESULTS The positive rate of VEGF-C mRNA in the lymph node metastasis tissues was 85.7% compared with negative controls (20%, P < .05). The density of lymphatic vessels in the metastasis group was 6.65 ± 1.57 compared with the negative group (3.75 ± 1.47, P < .05). Protein and mRNA of VEGF-C were reduced in transfected cells. Proliferation was inhibited as well. CONCLUSIONS VEGF-C can increase the invasiveness of gastric cancer and promote lymphangiogenesis in adjacent tissues. Transfection with antisense VEGF-C can reduce the expression of VEGF-C and inhibit the proliferation. VEGF-C can inhibit the tumor growth and reduce its metastasis and recurrence.
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Affiliation(s)
- Peng Zhu
- Department of Gastroenterological Surgery, Second Affiliated Hospital of Chongqing Medical University, 76 Linjiang Rd., Chongqing, People's Republic of China
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Sikkema AH, de Bont ESJM, Molema G, Dimberg A, Zwiers PJ, Diks SH, Hoving EW, Kamps WA, Peppelenbosch MP, den Dunnen WFA. Vascular endothelial growth factor receptor 2 (VEGFR-2) signalling activity in paediatric pilocytic astrocytoma is restricted to tumour endothelial cells. Neuropathol Appl Neurobiol 2011; 37:538-48. [PMID: 21208252 DOI: 10.1111/j.1365-2990.2011.01160.x] [Citation(s) in RCA: 21] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/22/2022]
Abstract
AIMS Tumours depend on angiogenesis for enhanced tumour cell survival and progression. Vascular endothelial growth factor receptor (VEGFR) signalling plays a major part in this process. Previously, we evaluated tyrosine kinase activity in paediatric brain tumour tissue lysates using a peptide microarray containing 144 different tyrosine kinase peptide substrates. When applied to paediatric pilocytic astrocytoma tissue, this analysis revealed extensive phosphorylation of VEGFR-derived peptides. The aim of the current study was to validate this result and determine the presence of VEGFR-2 activity in paediatric pilocytic astrocytoma as the main VEGFR in terms of mitogenic signalling. In addition, the localization of VEGFR1-3 mRNA expression was assessed. METHODS VEGFR-2 phosphorylation was determined by adopting a proximity ligation assay approach. Enrichment of endothelial markers and VEGFRs in tumour endothelium was determined by quantitative polymerase chain reaction (qPCR) analysis of laser-microdissected blood vessels. RESULTS Proximity ligation assays on tumour cryosections showed the presence of phosphorylation of VEGFR-2, which primarily localized to vascular endothelium. qPCR analysis of endothelial markers and VEGFRs showed a 13.6-fold average enrichment of VEGFR-2 expression in the laser-microdissected endothelium compared to whole tumour. Also the expression of VEGFR-1 and -3 was highly enriched in the endothelium fraction with an average fold-enrichment of 16.5 and 50.8 respectively. CONCLUSIONS Phosphorylated VEGFR-2 is detected on endothelial cells in paediatric pilocytic astrocytoma. Furthermore, endothelial cells are the main source of VEGFR1-3 mRNA expression. This suggests a crucial role for VEGF/VEGFR-induced angiogenesis in the progression and maintenance of these tumours.
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Affiliation(s)
- A H Sikkema
- Paediatric Oncology Division, Beatrix Children's Hospital, University of Groningen, Groningen, the Netherlands
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VEGF-C as a decision-making biomarker for selected patients with invasive bladder cancer who underwent bladder-preserving radical surgery. Arch Med Res 2011; 42:405-11. [PMID: 21821075 DOI: 10.1016/j.arcmed.2011.07.006] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/03/2011] [Accepted: 07/18/2011] [Indexed: 11/21/2022]
Abstract
BACKGROUND AND AIMS We proved the feasibility of radical transurethral resection in selected patients with muscle-invasive bladder cancer with a minimum follow-up of >5 years. A follow-up schedule was developed based on progression and recurrence during this period. METHODS The study included 93 patients with invasive bladder cancer treated by radical transurethral resection. Student t test was used for continuous variables to establish clinical progression predictive factors. VEGF-C protein expressions were tested by immunohistochemistry postsurgery. RESULTS The overall survival and disease-specific survival rates for all 93 patients were 59.1% and 65.2%, respectively. The clinical stage of the tumor influenced overall survival (p = 0.024) and disease-specific survival (p = 0.047). A significantly higher overall survival and disease-specific survival rate for patients with low levels of VEGF-C was 69.6% and 75.0%, respectively, than for those with high levels of VEGF-C (45.9 and 54.1%, respectively, p <0.05).The presence of bladder Tis reduced the survival rate (41.2 vs. 65.3%) and disease-specific survival (45.4 vs. 72.1%). Sensitivity, specificity and accuracy of VEGF-C in the evaluation of disease progression were 76.7, 77.8, 77.4%, respectively. CONCLUSIONS Patients with T2 stage, low level of VEGF-C and absence of bladder Tis were associated with high overall survival and disease-specific survival rate. VEGF-C level can evaluate disease progression and assist in choosing the appropriate treatment.
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Li Z, Qi F, Miao J, Zu X, He W, Wang L, Qi L. Vascular Endothelial Growth Factor-C Associated with Computed Tomography Used in the Diagnosis of Lymph Node Metastasis of Bladder Carcinoma. Arch Med Res 2010; 41:606-10. [DOI: 10.1016/j.arcmed.2010.11.003] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/23/2010] [Accepted: 10/06/2010] [Indexed: 11/16/2022]
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Tsirlis TD, Kostakis A, Papastratis G, Masselou K, Vlachos I, Papachristodoulou A, Nikiteas NI. Predictive significance of preoperative serum VEGF-C and VEGF-D, independently and combined with Ca19-9, for the presence of malignancy and lymph node metastasis in patients with gastric cancer. J Surg Oncol 2010; 102:699-703. [DOI: 10.1002/jso.21677] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/21/2022]
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Ran S, Volk L, Hall K, Flister MJ. Lymphangiogenesis and lymphatic metastasis in breast cancer. ACTA ACUST UNITED AC 2009; 17:229-51. [PMID: 20036110 DOI: 10.1016/j.pathophys.2009.11.003] [Citation(s) in RCA: 181] [Impact Index Per Article: 11.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/29/2009] [Revised: 04/11/2009] [Accepted: 10/23/2009] [Indexed: 01/03/2023]
Abstract
Lymphatic metastasis is the main prognostic factor for survival of patients with breast cancer and other epithelial malignancies. Mounting clinical and experimental data suggest that migration of tumor cells into the lymph nodes is greatly facilitated by lymphangiogenesis, a process that generates new lymphatic vessels from pre-existing lymphatics with the aid of circulating lymphatic endothelial progenitor cells. The key protein that induces lymphangiogenesis is vascular endothelial growth factor receptor-3 (VEGFR-3), which is activated by vascular endothelial growth factor-C and -D (VEGF-C and VEGF-D). These lymphangiogenic factors are commonly expressed in malignant, tumor-infiltrating and stromal cells, creating a favorable environment for generation of new lymphatic vessels. Clinical evidence demonstrates that increased lymphatic vessel density in and around tumors is associated with lymphatic metastasis and reduced patient survival. Recent evidence shows that breast cancers induce remodeling of the local lymphatic vessels and the regional lymphatic network in the sentinel and distal lymph nodes. These changes include an increase in number and diameter of tumor-draining lymphatic vessels. Consequently, lymph flow away from the tumor is increased, which significantly increases tumor cell metastasis to draining lymph nodes and may contribute to systemic spread. Collectively, recent advances in the biology of tumor-induced lymphangiogenesis suggest that chemical inhibitors of this process may be an attractive target for inhibiting tumor metastasis and cancer-related death. Nevertheless, this is a relatively new field of study and much remains to be established before the concept of tumor-induced lymphangiogenesis is accepted as a viable anti-metastatic target. This review summarizes the current concepts related to breast cancer lymphangiogenesis and lymphatic metastasis while highlighting controversies and unanswered questions.
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Affiliation(s)
- Sophia Ran
- Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University School of Medicine, 801 N. Rutledge, Springfield, IL 62794-9678, USA
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Schoppmann SF, Tamandl D, Roberts L, Jomrich G, Schoppmann A, Zwrtek R, Dubsky P, Gnant M, Jakesz R, Birner P. HER2/neu expression correlates with vascular endothelial growth factor-C and lymphangiogenesis in lymph node-positive breast cancer. Ann Oncol 2009; 21:955-60. [PMID: 19940005 DOI: 10.1093/annonc/mdp532] [Citation(s) in RCA: 45] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/01/2023] Open
Abstract
BACKGROUND Vascular endothelial growth factor-C (VEGF-C) is the main inducer of lymphangiogenesis. VEGF-C overexpression is associated with lymphovascular tumor cell invasion, an increased rate of lymph node metastasis and adverse prognosis in various human cancers. However, little is known about the upstream inducers of VEGF-C expression. Recent studies have shown that human epidermal growth factor receptor 2 (HER2/neu) overexpression is associated with high VEGF-C levels in human breast cancer cells. In addition to blocking of HER2/neu, tyrosine kinase significantly decreased VEGF-C expression in vitro. PATIENTS AND METHODS VEGF-C expression, lymphatic microvessel density (LMVD), lymphovascular invasion (LVI) and HER2/neu expression were evaluated with immunohistochemical/FISH methods in a collective of 150 lymph node-positive human breast cancers with long-term follow-up. RESULTS Cases with 3+ HER2/neu protein expression showed a significantly stronger VEGF-C expression than all others cases (P = 0.006). In addition, we found a significant correlation between VEGF-C expression and LMVD (P = 0.012) and a strong positive association between LMVD and LVI (P < 0.001). CONCLUSION Our data provide evidence for a clinically relevant association between HER2/neu and VEGF-C expression in human breast cancer. Inhibiting HER2/neu may reduce tumor progression by blocking VEGF-C-mediated tumor cell proliferation and lymphogenic metastasis.
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Affiliation(s)
- S F Schoppmann
- Department of Surgery, Medical University of Vienna, Vienna, Austria.
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Eccles S, Paon L, Sleeman J. Lymphatic metastasis in breast cancer: importance and new insights into cellular and molecular mechanisms. Clin Exp Metastasis 2007; 24:619-36. [PMID: 17985200 DOI: 10.1007/s10585-007-9123-5] [Citation(s) in RCA: 36] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/14/2007] [Accepted: 10/19/2007] [Indexed: 02/08/2023]
Abstract
Lymph node metastasis is the main prognosis factor in a number of malignancies, including breast carcinomas. The means by which lymph node metastases arise is not fully understood, and many questions remain about their importance in the further spread of breast cancer. Nevertheless, a number of key cellular and molecular mechanisms of lymphatic metastasis have been identified. These include induction of intra- or peri-tumoral lymphangiogenesis or co-option of existing lymphatic vessels to allow tumour cells to enter the lymphatics, although it remains to be established whether this is primarily an active or passive process. Gene expression microarrays and functional studies in vitro and in vivo, together with detailed clinical observations have identified a number of molecules that can play a role in the genesis of lymph node metastases. These include the well-recognised lymphangiogenic cytokines VEGF-C and VEGF-D as well as chemokine-receptor interactions, integrins and downstream signalling pathways. This paper briefly reviews current clinical and experimental evidence for the underlying mechanisms and significance of lymphatic metastasis in breast cancer and highlights questions that still need to be addressed.
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Affiliation(s)
- Suzanne Eccles
- Cancer Research UK Centre for Cancer Therapeutics, McElwain Laboratories, The Institute of Cancer Research, Cotswold Road, Sutton, Surrey, SM2 5NG, UK.
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Zhou HC, Kang CY, Zhang Y, Li JC. Construction and expression of small interfering RNA vector targeting on vascular endothelial growth factor-C. Shijie Huaren Xiaohua Zazhi 2007; 15:1549-1553. [DOI: 10.11569/wcjd.v15.i13.1549] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: To clone the recombinant plasmid affecting vascular endothelial growth factor-C (VEGF-C) gene transcription by RNA interference and observe VEGF-C expression in human gastric cancer cell.
METHODS: Two DNA sequences containing small hairpin structure were designed and synthesized. The complement form was obtained by annealing and then cloned into vector pSilencer3.1-H1. The recombinant plasmid was transformed into strain DH5α. After amplification and purification, the target plasmid, identified by restriction enzyme, was obtained sequenced. Finally, gastric carcinoma cell line SGC-7901 was transfected with the plasmid, and the protein expression of VEGF-C was detected by Western blot.
RESULTS: After enzyme digestion and sequencing, the VEGF-C siRNA expression vector was successfully constructed. Western blot analysis showed that the protein expression of VEGF-C was markedly decreased after transfection, especially in the cells transfected with pSilencer3.1-VEGF-C1, and the inhibitory rate was 81.2%, significantly different from that in the negative control group (P < 0.05).
CONCLUSION: The recombinant plasmid containing VEGF-C siRNA is successfully constructed. RNAi may continually and stably suppress VEGF-C protein expression, which is a potential approach for tumor gene therapy.
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Mohammed RAA, Green A, El-Shikh S, Paish EC, Ellis IO, Martin SG. Prognostic significance of vascular endothelial cell growth factors -A, -C and -D in breast cancer and their relationship with angio- and lymphangiogenesis. Br J Cancer 2007; 96:1092-100. [PMID: 17353919 PMCID: PMC2360132 DOI: 10.1038/sj.bjc.6603678] [Citation(s) in RCA: 140] [Impact Index Per Article: 7.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/02/2022] Open
Abstract
Vascular endothelial cell growth factors (VEGF)-A, -C and -D have potent angio and lymphangiogenic functions in experimental models, although their role in the progression of human breast cancer is unclear. The aims of the current study were to examine the relationship between the expression of the aforementioned growth factors with the angio and lymphangiogenic characteristics of breast cancer, and to assess their suitability as potential prognostic factors. Paraffin-embedded sections of 177 primary invasive breast cancer, with complete clinical follow-up information for 10 years, were stained for VEGF-A, -C, -D, podoplanin and CD34 using standard immunohistochemical approaches. The expression of the growth factors was correlated with clinicopathological criteria and patients’ survival. Lymph vessel density (LVD) and microvessel density (MVD) were assessed and correlated with expression of the growth factors. Vascular endothelial cell growth factor-A, -C and -D were highly expressed in 40, 37 and 42% of specimens, respectively. High expression of VEGF-A and - C, but not of -D, was associated with a higher LVD (P=0.013 and P=0.014, respectively), a higher MVD (P<0.001 and P=0.002, respectively), the presence of lymph node metastasis (P<0.001 and P<0.001, respectively), distant metastasis (P=0.010 and P=0.008, respectively) and a shorter Overall Survival (P=0.029 and 0.028, respectively). In conclusion, breast cancers that express high levels of VEGF-A and -C are characterised by a poor prognosis, likely through the induction of angio and lymphangiogenesis. Examination of expression of VEGF-A and -C in breast cancer may be beneficial in the identification of a subset of tumours that have a higher probability of recurrence and metastatic spread.
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Affiliation(s)
- R A A Mohammed
- Department of Clinical Oncology, University Hospitals, City Hospital Campus, University of Nottingham, Hucknall Road, NG5 1PB, Nottingham, UK
- Histopathology Departments, University Hospitals, City Hospital Campus, University of Nottingham, Hucknall Road, NG5 1PB, Nottingham, UK
| | - A Green
- Histopathology Departments, University Hospitals, City Hospital Campus, University of Nottingham, Hucknall Road, NG5 1PB, Nottingham, UK
| | - S El-Shikh
- Histopathology Departments, University Hospitals, City Hospital Campus, University of Nottingham, Hucknall Road, NG5 1PB, Nottingham, UK
| | - E C Paish
- Histopathology Departments, University Hospitals, City Hospital Campus, University of Nottingham, Hucknall Road, NG5 1PB, Nottingham, UK
| | - I O Ellis
- Histopathology Departments, University Hospitals, City Hospital Campus, University of Nottingham, Hucknall Road, NG5 1PB, Nottingham, UK
| | - S G Martin
- Department of Clinical Oncology, University Hospitals, City Hospital Campus, University of Nottingham, Hucknall Road, NG5 1PB, Nottingham, UK
- E-mail:
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Su JL, Yen CJ, Chen PS, Chuang SE, Hong CC, Kuo IH, Chen HY, Hung MC, Kuo ML. The role of the VEGF-C/VEGFR-3 axis in cancer progression. Br J Cancer 2006; 96:541-5. [PMID: 17164762 PMCID: PMC2360045 DOI: 10.1038/sj.bjc.6603487] [Citation(s) in RCA: 201] [Impact Index Per Article: 10.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/06/2023] Open
Abstract
Vascular endothelial growth factor (VEGF) receptor 3 (VEGFR-3) (also called VEGFR-3) is activated by its specific ligand, VEGF-C, which promotes cancer progression. The VEGF-C/VEGFR-3 axis is expressed not only by lymphatic endothelial cells but also by a variety of human tumour cells. Activation of the VEGF-C/VEGFR-3 axis in lymphatic endothelial cells can facilitate metastasis by increasing the formation of lymphatic vessels (lymphangiogenesis) within and around tumours. The VEGF-C/VEGFR-3 axis plays a critical role in leukaemic cell proliferation, survival, and resistance to chemotherapy. Moreover, activation of the VEGF-C/VEGFR-3 axis in several types of solid tumours enhances cancer cell mobility and invasion capabilities, promoting cancer cell metastasis. In this review, we discuss the novel function and molecular mechanism of the VEGF-C/VEGFR-3 axis in cancer progression.
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Affiliation(s)
- J-L Su
- Institute of Medical Science, College of Medicine, China Medical University, Taichung 404, Taiwan
- Center for Molecular Medical, China Medical University Hospital, Taichung 404, Taiwan
- Department of Biotechnology and Bioinformatics, Asia University, Taichung 41354, Taiwan
- Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA
| | - C-J Yen
- Department of Internal Medicine, National Cheng Kung University Hospital, Tainan 70428, Taiwan
- Institute of Clinical Medicine, National Cheng Kung University Hospital, Tainan 70428, Taiwan
| | - P-S Chen
- Institute of Toxicology, College of Medicine, National Taiwan University, Taipei 100, Taiwan
| | - S-E Chuang
- Division of Cancer Research, National Health Research Institutes, Taipei 10016, Taiwan
| | - C-C Hong
- Division of Cancer Research, National Health Research Institutes, Taipei 10016, Taiwan
| | - I-H Kuo
- Institute of Toxicology, College of Medicine, National Taiwan University, Taipei 100, Taiwan
| | - H-Y Chen
- Institute of Toxicology, College of Medicine, National Taiwan University, Taipei 100, Taiwan
| | - M-C Hung
- Department of Molecular and Cellular Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA
- Graduate School of Biomedical Sciences, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA
| | - M-L Kuo
- Institute of Toxicology, College of Medicine, National Taiwan University, Taipei 100, Taiwan
- Angiogenesis Research Center, National Taiwan University, Taipei, Taiwan
- Laboratory of Molecular and Cellular Toxicology, Institute of Toxicology, College of Medicine, National Taiwan University, No. 1, Section 1, Jen-Ai Road, Taipei, Taiwan. E-mail:
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Grau SJ, Trillsch F, Herms J, Thon N, Nelson PJ, Tonn JC, Goldbrunner R. Expression of VEGFR3 in glioma endothelium correlates with tumor grade. J Neurooncol 2006; 82:141-50. [PMID: 17115285 DOI: 10.1007/s11060-006-9272-4] [Citation(s) in RCA: 45] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/17/2006] [Accepted: 09/13/2006] [Indexed: 11/27/2022]
Abstract
Angiogenic processes are regulated by vascular endothelial growth factors (VEGFs) and their receptors VEGFR1 (Flt-1), 2 (Flk-1) and 3 (Flt-4). While VEGFR2 is thought to play a central role in tumor angiogenesis, anti-angiogenic therapies targeting VEGFR2 in glioma models can show escape phenomena with secondary onset of angiogenesis. The purpose of this study was to find explanations for these processes by searching for alternative pathways regulating glioma angiogenesis and reveal a correlation with tumor grade. Thus, VEGFR3, which is not expressed in normal brain, and its ligands VEGF-C and -D, were assessed in high grade (WHO degrees IV, glioblastomas, GBM) and low grade gliomas [WHO degrees II astrocytomas (AII)]. In all GBM, a strong protein expression of VEGFR3 was found on tumor endothelium, VEGF-C and -D expression was found on numerous cells in areas of high vascularization. On RNA level, a significant up-regulation of VEGFR3 was detected in GBM compared to AII and non-neoplastic brain. In AII, only very moderate VEGFR3, VEGF-C and -D expression was found on protein and RNA level indicating a correlation of VEGFR3 expression with tumor grade. VEGFR3 signal in both grades was found predominantly on endothelial cells, confirmed by VEGFR3 expression on isolated CD31 positive cells and the expression of various endothelial markers on VEGFR3-positive cells isolated from GBM. The demonstration of a complete angiogenic signaling system that is dependent on tumor grade may influence the traditional paradigm of glioma angiogenesis and may provide a basis for more effective anti-angiogenic treatment strategies.
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Affiliation(s)
- S J Grau
- Department of Neurosurgery, Klinikum Grosshadern, Ludwig-Maximilians-University Munich, Marchioninistr. 15, 81373 , Munich, Germany,
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Zhang J, Ji J, Yuan F, Zhu L, Yan C, Yu YY, Liu BY, Zhu ZG, Lin YZ. Cyclooxygenase-2 expression is associated with VEGF-C and lymph node metastases in gastric cancer patients. Biomed Pharmacother 2006; 59 Suppl 2:S285-8. [PMID: 16507394 DOI: 10.1016/s0753-3322(05)80047-2] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/01/2023] Open
Abstract
Previous studies have suggested that cyclooxygenase-2 (COX-2) over-expression is associated with angiogenesis in gastric cancer. However, the relationship between COX-2 and lymphangiogenesis is still unclear. The aim of this study was to determine the relationship between COX-2 expression and lymphangiogenic factor, vascular endothelial growth factor-C (VEGF-C), in human gastric cancer, as well as to correlate with clinicopathological parameters. Sixty-three gastric cancer patients underwent radical gastrectomy (D2 or D3) were enrolled in this study. The expression of COX-2 and VEGF-C were detected by immunohistochemistry, and the small lymphatic vessels were immunohistochemically stained by LYVE-1 antibody. The association between COX-2 and VEGF-C expressions and clinicopathological parameters (such as gender, tumor location, lymph node status and Lauren classification) were determined. VEGF-C over-expression was observed in 33 of 63 patients (52%), while COX-2 over-expression occurred in 42 of 63 tumor samples (67%). Presence of microlymphatic vessels with LYVE-1 staining was found in 35 cases. COX-2 over-expression was highly correlated with VEGF-C over-expression (P = 0.032), microlymphatic vessels (P = 0.002) as well as presence of metastatic lymph nodes (P = 0.007). However, no significant correlation was found between COX-2 expression and other clinicopathological parameters. Our data suggest that COX-2 expression is associated with lymphangiogenesis and lymph node metastasis in human gastric carcinoma. This raises the possibility that COX-2-mediated VEGF-C over-expression might promote lymph node metastasis via lymphangiogenesis pathway in patients with gastric cancer.
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Affiliation(s)
- J Zhang
- Department of Surgery, Rui Jin Hospital, Shanghai Institute of Digestive Surgery, Shanghai Second Medical University, China
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Thiele W, Sleeman JP. Tumor-induced lymphangiogenesis: a target for cancer therapy? J Biotechnol 2006; 124:224-41. [PMID: 16497404 DOI: 10.1016/j.jbiotec.2006.01.007] [Citation(s) in RCA: 61] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/21/2005] [Revised: 11/07/2005] [Accepted: 01/04/2006] [Indexed: 12/16/2022]
Abstract
Recent advances in understanding the biology of lymphangiogenesis, the new growth of lymphatic vessels, have cast new light on the molecular basis of metastasis to regional lymph nodes. The receptor tyrosine kinase VEGFR-3 is virtually exclusively expressed on lymphatic but not blood endothelium in the adult, and activation of VEGFR-3 by its ligands VEGF-C and VEGF-D is sufficient to induce lymphangiogenesis. Correlative studies with human tumors and functional studies using animal tumor models show that increased levels of VEGF-C or VEGF-D in tumors lead to enhanced numbers of lymphatic vessels in the vicinity of tumors, which in turn promotes metastasis to regional lymph nodes by providing a greater number of entry sites into the lymphatic system for invading tumor cells. These findings have prompted studies to investigate whether inhibitors of VEGFR-3 activation might represent novel therapeutic agents for the suppression of metastasis. However, a number of points regarding the therapeutic potential of anti-lymphangiogenic treatments in the context of cancer remain to be addressed. The spectrum and relative importance of molecules that induce lymphangiogenesis and the regulation of their expression during tumor progression, the reversibility of tumor-induced lymphangiogenesis, and possible side-effects of anti-lymphangiogenesis-based therapies all need to be investigated. Most importantly, the extent to which lymph node metastases contribute to the formation of metastases in other organs remains to be elucidated. These aspects are the focus of this review, and their investigation should serve as a roadmap to possible translational application.
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Affiliation(s)
- Wilko Thiele
- Forschungszentrum Karlsruhe, Institut für Toxikologie und Genetik, Germany
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31
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Yonemura Y, Endo Y, Tabata K, Kawamura T, Yun HY, Bandou E, Sasaki T, Miura M. Role of VEGF-C and VEGF-D in lymphangiogenesis in gastric cancer. Int J Clin Oncol 2006; 10:318-27. [PMID: 16247658 DOI: 10.1007/s10147-005-0508-7] [Citation(s) in RCA: 51] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/03/2005] [Accepted: 05/20/2005] [Indexed: 01/30/2023]
Abstract
BACKGROUND The molecular mechanisms of lymphangiogenesis induced by vascular endothelial growth factor (VEGF)-C and VEGF-D in gastric cancer were studied. METHODS VEGF-C and VEGF-D gene expression vectors were transfected into the gastric cancer cell line KKLS, which did not originally express VEGF-C and VEGF-D, and stable transfectants (KKLS/VEGF-C and KKLS/VEGF-D) were established. The cell lines were inoculated into the subserosal layer of the stomach and subcutaneous tissue of nude mice. RESULTS VEGF-C and VEGF-D expression in KKLS/VEGF-C and KKLS/VEGF-D cells was found by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. Expression of mouse VEGF receptor (VEGFR)-2 and mouse VEGFR-3 mRNA was detected in the KKLS/VEGF-C and KKLS/VEGF-D gastric tumors. Newly formed lymphatic vessels were detected not only in the periphery but also in the center of the tumors. The intratumor lymphatic vessels connected with the preexisting lymphatic vessels in the muscularis mucosa. The average numbers of lymphatic vessels in KKLS/VEGF-C (52.0 +/- 9.5) and KKLS/VEGF-D (16.4 +/- 0.6) gastric tumors were significantly higher than that in the KKLS/control vector tumors (4.0 +/- 1.4). CONCLUSION VEGF-C and VEGF-D may induce neoformation of lymphatic vessels in experimental gastric tumors by the induction of VEGFR-3 expression.
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Affiliation(s)
- Yutaka Yonemura
- Gastric Surgery Division, Shizuoka Cancer Center, 1007 Shimo-Nagakubo, Shizuoka 411-8777, Japan.
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Chen Z, Varney ML, Backora MW, Cowan K, Solheim JC, Talmadge JE, Singh RK. Down-regulation of vascular endothelial cell growth factor-C expression using small interfering RNA vectors in mammary tumors inhibits tumor lymphangiogenesis and spontaneous metastasis and enhances survival. Cancer Res 2005; 65:9004-11. [PMID: 16204074 DOI: 10.1158/0008-5472.can-05-0885] [Citation(s) in RCA: 105] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
Abstract
Tumor production of vascular endothelial cell growth factor (VEGF)-C is associated with tumor lymphangiogenesis and lymph node metastasis. In this study, we examined the effects of small interfering RNA (siRNA)-mediated inhibition of VEGF-C on murine mammary tumor growth, metastasis, and survival. The mRNA and protein expression of VEGF-C in murine mammary tumor cells stably transfected with a VEGF-C siRNA vector were significantly lower compared with VEGF-C-control vector-transfected cells. Cl66-siVEGFC tumors had lower levels of lymphangiogenesis and lymph node and spontaneous lung metastasis than Cl66-control tumors. However, we did not observe significant differences in primary tumor growth and experimental lung metastasis between mice injected with Cl66-siVEGFC and Cl66-control cells. In addition, mice bearing Cl66-siVEGFC tumors lived significantly longer than mice bearing Cl66-control tumors. Furthermore, our data suggest that inhibition of VEGF-C modulates immune cell infiltration and their function, which might be critical in tumor immunity. In summary, our data show that inhibition of VEGF-C expression using siRNA-mediated gene silencing vectors reduces lymphangiogenesis and lymph node and spontaneous lung metastasis, and enhances survival.
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Affiliation(s)
- Zhengtang Chen
- Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska 68198-5845, USA
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Weigelt B, Wessels LFA, Bosma AJ, Glas AM, Nuyten DSA, He YD, Dai H, Peterse JL, van't Veer LJ. No common denominator for breast cancer lymph node metastasis. Br J Cancer 2005; 93:924-32. [PMID: 16189523 PMCID: PMC2361648 DOI: 10.1038/sj.bjc.6602794] [Citation(s) in RCA: 70] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/30/2022] Open
Abstract
The axillary lymph node status is the most powerful prognostic factor for breast cancer patients to date. The molecular mechanisms that control lymph node metastasis, however, remain poorly understood. To define patterns of genes or gene regulatory pathways that drive breast cancer lymph node metastasis, we compared the gene expression profiles of 15 primary breast carcinomas and their matching lymph node metastases using microarrays. In general, primary breast carcinomas and lymph node metastases do not differ at the transcriptional level by a common subset of genes. No classifier or single gene discriminating the group of primary tumours from those of the lymph node metastases could be identified. Also, in a series of 295 breast tumours, no classifier predicting lymph node metastasis could be developed. However, subtle differences in the expression of genes involved in extracellular-matrix organisation and growth factor signalling are detected in individual pairs of matching primary and metastatic tumours. Surprisingly, however, different sets of these genes are either up- or downregulated in lymph node metastases. Our data suggest that breast carcinomas do not use a shared gene set to accomplish lymph node metastasis.
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Affiliation(s)
- B Weigelt
- Division of Experimental Therapy, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands
| | - L F A Wessels
- Division of Diagnostic Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands
- Information and Communication Theory Group, Delft University of Technology, 2600 GA Delft, The Netherlands
| | - A J Bosma
- Division of Experimental Therapy, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands
| | - A M Glas
- Division of Diagnostic Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands
| | - D S A Nuyten
- Division of Radiotherapy, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands
| | - Y D He
- Rosetta Inpharmatics LLC, Seattle, WA 98109, USA
| | - H Dai
- Rosetta Inpharmatics LLC, Seattle, WA 98109, USA
| | - J L Peterse
- Division of Diagnostic Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands
| | - L J van't Veer
- Division of Experimental Therapy, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands
- Division of Diagnostic Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands
- The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands. E-mail: Division of Experimental Therapy, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands,
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Van Trappen PO, Pepper MS. Lymphangiogenesis in human gynaecological cancers. Angiogenesis 2005; 8:137-45. [PMID: 16211357 DOI: 10.1007/s10456-005-9008-7] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/22/2004] [Accepted: 02/25/2005] [Indexed: 01/23/2023]
Abstract
The metastatic spread of tumor cells is responsible for the majority of cancer deaths, and with few exceptions, all cancers can metastasize. Clinical findings have for a long time suggested that by providing a pathway for tumor cell dissemination, tumor-associated lymphatics act as key components of metastatic spread. This is believed to occur principally via pre-existing and possibly also newly formed lymphatics (lymphangiogenesis). Increased expression of vascular endothelial growth factor-C (VEGF-C) and VEGF-D in primary tumors correlates with increased dissemination of tumor cells to regional lymph nodes (LNs) in a variety of human carcinomas. Here we will review the mechanisms of lymphangiogenesis, particularly in the context of metastatic tumor spread, and will critically examine the role of VEGF-C and VEGF-D in this process in gynaecological cancers. Potential anti-lymphangiogenic strategies are also discussed.
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Affiliation(s)
- Philippe O Van Trappen
- Gynaecological Cancer Centre and Cancer Research UK Translational Oncology Laboratory, Queen Mary University of London, St Bartholomew's Hospital, London, UK
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Abstract
This review advances the hypothesis that the function of vascular endothelial growth factor (VEGF) in breast cancer is not limited to angiogenesis, and that VEGF signaling in breast carcinoma cells is important for the ability of these cells to evade apoptosis and progress towards invasive and metastatic disease. In other terms, VEGF signaling provides a selective advantage for the survival and dissemination of breast carcinoma cells that may be independent of angiogenesis. The key component of this hypothesis is that breast carcinoma cells express specific VEGF receptors and that these receptors respond to autocrine VEGF, resulting in the activation of signaling pathways that impede apoptosis and promote cell migration. A related hypothesis, which is developed in this review, is that the alpha6beta4 integrin, which has been implicated in the survival and motility of breast cancer cells, can stimulate the translation of VEGF mRNA and, consequently, autocrine VEGF signaling. These findings imply that VEGF and VEGF receptor-based therapeutics, in addition to targeting angiogenesis, may also target tumor cells directly.
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Affiliation(s)
- Arthur M Mercurio
- Department of Cancer Biology and the Cancer Center, University of Massachusetts Medical School, Worcester, MA 01605, USA.
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Affiliation(s)
- Marc G Achen
- Ludwig Institute for Cancer Research, Royal Melbourne Hospital, Victoria, Australia.
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Kaushal V, Mukunyadzi P, Dennis RA, Siegel ER, Johnson DE, Kohli M. Stage-Specific Characterization of the Vascular Endothelial Growth Factor Axis in Prostate Cancer: Expression of Lymphangiogenic Markers Is Associated with Advanced-Stage Disease. Clin Cancer Res 2005. [DOI: 10.1158/1078-0432.584.11.2] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
Abstract
Abstract
Purpose: The vascular endothelial growth factor (VEGF) family plays a critical role in tumor angiogenesis and lymphangiogenesis. We characterized, at the mRNA and protein levels, the expression of VEGF-A and VEGF-D and their cognate receptors, VEGFR-1, VEGFR-2, and VEGFR-3 in early- and advanced-stage prostate cancer specimens.
Experimental Design: The levels of VEGF-A and VEGF-D mRNA in early- and advanced-stage specimens were compared using an angiogenic gene array and were confirmed by quantitative real-time PCR. Receptor protein levels and activation status were determined by immunoblotting. Spatial expression of the proteins was evaluated using immunohistochemistry with fresh and archival tissues from benign prostatic hypertrophy specimens, early-stage prostate specimens, and advanced-stage metastatic specimens. Circulating plasma levels of these growth factors were measured using ELISAs.
Results: We observed that expression patterns of VEGF isotypes corresponded to the prostate cancer stage: high expression of angiogenic growth factor VEGF-A was observed in early-stage prostate specimens, whereas high expression of lymphangiogenic growth factor VEGF-D was associated with advanced-stage metastatic disease. All VEGF receptors were present at variable levels in all specimens, but their activation states varied in a stage-specific manner. VEGFR-1 and, to a limited extent, VEGFR-2 were activated in early-stage specimens, whereas VEGFR-2 and VEGFR-3 were activated in advanced-stage specimens.
Conclusions: Our results suggest that lymphangiogenic markers, such as VEGF-D and VEGFR-2 and VEGFR-3, may be better than angiogenic markers as targets of therapeutic intervention in advanced-stage prostate disease.
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Affiliation(s)
| | | | | | - Eric R. Siegel
- 4Biostatistics, University of Arkansas for Medical Sciences and Departments of
| | | | - Manish Kohli
- 1Internal Medicine, Departments of
- 6Hematology/Oncology, Central Arkansas Veterans Healthcare System, Little Rock, Arkansas
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Jia YT, Li ZX, He YT, Liang W, Yang HC, Ma HJ. Expression of vascular endothelial growth factor-C and the relationship between lymphangiogenesis and lymphatic metastasis in colorectal cancer. World J Gastroenterol 2004; 10:3261-3. [PMID: 15484296 PMCID: PMC4572291 DOI: 10.3748/wjg.v10.i22.3261] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/15/2022] Open
Abstract
AIM: To investigate the expression of vascular endothelial growth factor-C (VEGF-C) and the relationship between VEGF-C and lymphangiogenesis, lymph node metastasis in colorectal cancer.
METHODS: Fifty six cases of colorectal cancer were selected randomly. Expression of VEGF-C was detected by immuno- histochemistry, and lymphatic vessels were stained by enzyme histochemical method.
RESULTS: VEGF-C expression was found in 66.7% (37/56) patients. In VEGF-C positive and negative patients, the lymphatic vessel density was 25.16 ± 7.52 and 17.14 ± 7.22, respectively (P < 0.05). The rate of lymph node metastasis in VEGF-C positive patients (81.1%) was significantly higher than that in the negative group (42.1%).
CONCLUSION: VEGF-C expression may induce lymphangiogenesis in colorectal cancer, as a result, tumor cells can entry the lymphatic vessels easily. VEGF-C may serve as a useful prognotic factor in colorectal carcinoma.
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Affiliation(s)
- Yi-Tao Jia
- Department of Oncology, the People's Hospital of Hebei Province, Shijiazhuang 050051, Hebei Province, China
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