• Breast cancer
• CCN5
• Ductal carcinoma in situ
• Estrogen receptor
• Invasive carcinoma

• Humans
• Female
• Breast Neoplasms/pathology
• Breast Neoplasms/metabolism
• Breast Neoplasms/genetics
• Receptors, Estrogen/metabolism
• Middle Aged
• Adult
• CCN Intercellular Signaling Proteins/metabolism
• Carcinoma, Intraductal, Noninfiltrating/pathology
• Carcinoma, Intraductal, Noninfiltrating/metabolism
• Aged
• Biomarkers, Tumor/metabolism
• Carcinoma, Ductal, Breast/pathology
• Carcinoma, Ductal, Breast/metabolism
• Immunohistochemistry
• Nephroblastoma Overexpressed Protein/metabolism
• Receptor, ErbB-2/metabolism
• Clinical Relevance
• Repressor Proteins

• CCN5
• E-cadherin
• breast cancer
• metastases
• primary lesion
• recurrent lesion

• macular degeneration
• retinal diseases

• Epithelial-Mesenchymal Transition/drug effects
• Retinal Pigment Epithelium/metabolism
• Retinal Pigment Epithelium/drug effects
• Retinal Pigment Epithelium/pathology
• Animals
• Humans
• Mice
• Vascular Endothelial Growth Factor A/metabolism
• Macular Degeneration/metabolism
• Macular Degeneration/pathology
• Macular Degeneration/drug therapy
• Macular Degeneration/chemically induced
• Cell Line
• Bevacizumab/pharmacology
• CCN Intercellular Signaling Proteins/metabolism
• CCN Intercellular Signaling Proteins/genetics
• Angiogenesis Inhibitors/pharmacology
• Ranibizumab/pharmacology
• Recombinant Fusion Proteins/pharmacology
• Signal Transduction/drug effects
• Repressor Proteins
• Receptors, Vascular Endothelial Growth Factor

• 2022R1A2C1004256 National Research Foundation of Korea

• CCN
• Extracellular matrix
• Matricellular protein
• Neoplastic tissue
• Tumor microenvironment

• I01 BX001002 BLRD VA
• I01 BX004497 BLRD VA
• IK6 BX005769 BLRD VA
• 1IK6BX005769-01 VA Research Career Scientist Award
• 5I01BX004497-03 VA Merit grant
• I01 BX001989 BLRD VA
• 5I01BX001002 VA Merit Grant

Breast cancer is the most frequent malignancy in women worldwide. Advanced breast cancer with distant organ metastases is considered incurable with currently available therapies. The vasculogenic mimicry (VM) process is associated with an invasive and metastatic cancer phenotype and a poor prognosis for human breast cancer patients. Our aim was to study the effect of WISP2, a matricellular protein, on VM. We found that WISP2 inhibits VM through inhibition of CYR61 protein expression and YAP-TAZ signaling. Our finding may open promising candidates for blocking VM in breast cancer.

Vasculogenic mimicry (VM) formed by aggressive tumor cells to create vascular networks connected with the endothelial cells, plays an important role in breast cancer progression. WISP2 has been considered as a tumor suppressor protein; however, the relationship between WISP2 and VM formation remains unclear. We used the in vitro tube formation assay and in vivo immunohistochemical analysis in a mouse model, and human breast tumors were used to evaluate the effect of WISP2 on VM formation. Here we report that WISP2 acts as a potent inhibitor of VM formation in breast cancer. Enforced expression of WISP2 decreased network formation while knockdown of WISP2 increased VM. Mechanistically, WISP2 increased retention of oncogenic activators YAP/TAZ in cytoplasm, leading to decreased expression of the angiogenic factor CYR61. Studies using an in vivo mouse model and human breast tumors confirmed the in vitro cell lines data. In conclusion, our results indicate that WISP2 may play a critical role in VM and highlight the critical role of WISP2 as a tumor suppressor.

• CYR61
• WISP2/CCN5
• YAP-TAZ signaling
• breast cancer
• vasculogenic mimicry

• CCN5
• EGCG
• FA-PEG-NPs
• PCNA
• TNBC
• bioavailability
• breast cancer
• drug delivery
• folic acid
• nanoparticles

• Animals
• Apoptosis/drug effects
• CCN Intercellular Signaling Proteins/agonists
• CCN Intercellular Signaling Proteins/metabolism
• Catechin/administration & dosage
• Catechin/analogs & derivatives
• Cell Line, Tumor
• Cell Survival/drug effects
• Female
• Humans
• Mice
• Nanoparticle Drug Delivery System
• Repressor Proteins/agonists
• Repressor Proteins/metabolism
• Spheroids, Cellular
• Triple Negative Breast Neoplasms/drug therapy
• Triple Negative Breast Neoplasms/pathology
• Xenograft Model Antitumor Assays

• I01 BX001002 BLRD VA
• P20 GM109024 NIGMS NIH HHS
• 1P20 GM109024 NIGMS NIH HHS
• 5I01BX001989-04 VA
• I01 BX004497 BLRD VA
• IK6 BX005769 BLRD VA
• U54 HD090216 NICHD NIH HHS
• I01BX001002-05 VA
• I01 BX001989 BLRD VA
• Center for Scientific Review

The crosstalk between tumor cells and the tumor microenvironment (TME), triggers a variety of critical signaling pathways and promotes the malignant progression of cancer. The success rate of cancer therapy through targeting single molecule of this crosstalk may be extremely low, whereas co-targeting multiple components could be complicated design and likely to have more side effects. The six members of cellular communication network (CCN) family proteins are scaffolding proteins that may govern the TME, and several studies have shown targeted therapy of CCN family proteins may be effective for the treatment of cancer. CCN protein family shares similar structures, and they mutually reinforce and neutralize each other to serve various roles that are tightly regulated in a spatiotemporal manner by the TME. Here, we review the current knowledge on the structures and roles of CCN proteins in different types of cancer. We also analyze CCN mRNA expression, and reasons for its diverse relationship to prognosis in different cancers. In this review, we conclude that the discrepant functions of CCN proteins in different types of cancer are attributed to diverse TME and CCN truncated isoforms, and speculate that targeting CCN proteins to rebalance the TME could be a potent anti-cancer strategy.

• CCN proteins
• isoforms
• pan-cancer
• targeted therapy
• tumor microenvironment

• epithelial–mesenchymal transition
• extracellular vesicles
• hypoxia
• pulsed-SILAC
• quantitative proteomics
• tumorigenesis
• tumour microenvironment

• MR/R008302/1 Medical Research Council

• CCN5
• PI3K/AKT signaling
• oral squamous cell carcinoma
• tumor suppressor

• Cancer
• Immunogenicity
• Nanoparticles
• PEGylation
• Polyethylene glycol

• 5I01BX001989-04 VA Merit Award
• I01 BX001002 BLRD VA
• I01 BX006279 BLRD VA
• I01BX001002-05 VA Merit Award
• I01 BX004497 BLRD VA
• IK6 BX005769 BLRD VA

Emerging evidence has demonstrated that WISP2 is critically involved in cell proliferation, migration, invasion and metastasis in cancers. However, the function of WISP2 in esophageal squamous cell carcinoma (ESCC) is largely unclear. Therefore, we aim to explore the effects and the potential mechanism of WISP2 on proliferation and motility and invasion of ESCC cells.

Cell proliferation was detected by MTT assay and apoptosis was measured by FACS in ESCC cells after WISP2 downregulation and overexpression. Cell migration and invasion were analyzed by wound healing assay and transwell migration assay, respectively. The expression of ERK-1/2, Slug and E-cadherin was measured by Western blot respectively. IHC was performed to measure the expression of WISP2 in ESCC tissues.

WISP2 overexpression is associated with survival in ESCC patients. WISP2 overexpression inhibited cell growth and induced cell apoptosis, suppressed cell migration and invasion in ESCC cells. Moreover, WISP overexpression retarded tumor growth in mouse model. WISP2 downregulation enhanced cell growth, inhibited apoptosis, promoted cell migration and invasion in ESCC cells. Mechanistically, WISP2 exerts its tumor suppressive functions via regulation of ERK1/2, Slug, and E-cadherin in ESCC cells.

Our findings suggest that activation of WISP2 could be a useful therapeutic strategy for the treatment of ESCC.

• Esophageal cancer
• Invasion
• Migration
• Proliferation
• WISP2

• Cancer
• Drug
• Oncogene
• Targets
• Therapy
• WISP

• CCN Intercellular Signaling Proteins/metabolism
• Carcinogenesis/metabolism
• Humans
• Neoplasms/metabolism
• Neoplasms/pathology
• Neoplasms/therapy
• Signal Transduction

• 81572936 National Natural Science Foundation of China
• 81773186 National Natural Science Foundation of China

Mouse models of breast cancer with specific molecular subtypes (e.g., ER or HER2 positive) in an immunocompetent or an immunocompromised environment significantly contribute to our understanding of cancer biology, despite some limitations, and they give insight into targeted therapies. However, an ideal triple-negative breast cancer (TNBC) mouse model is lacking. What has been missing in the TNBC mouse model is a sequential progression of the disease in an essential native microenvironment. This notion inspired us to develop a TNBC-model in syngeneic mice using a mammary intraductal (MIND) method. To achieve this goal, Mvt-1and 4T1 TNBC mouse cell lines were injected into the mammary ducts via nipples of FVB/N mice and BALB/c wild-type immunocompetent mice, respectively. We established that the TNBC-MIND model in syngeneic mice could epitomize all breast cancer progression stages and metastasis into the lungs via lymphatic or hematogenous dissemination within four weeks. Collectively, the syngeneic mouse-TNBC-MIND model may serve as a unique platform for further investigation of the underlying mechanisms of TNBC growth and therapies.

• Animals
• Cell Line, Tumor
• Cell Proliferation
• Cell Transformation, Neoplastic
• Disease Models, Animal
• Disease Progression
• Female
• Humans
• Lung Neoplasms/secondary
• Mammary Neoplasms, Experimental/pathology
• Mesoderm/pathology
• Mice
• Neoplasm Invasiveness
• Triple Negative Breast Neoplasms/pathology

• I01 BX001002 BLRD VA
• I01 BX001989 BLRD VA
• R01 CA207445 NCI NIH HHS
• VA Merit Award Grant
• Grace Hortense Greenley Trust, directed by The Research Foundation in memory of Eva Lee Caldwell
• Grace Hortense Greenley Trust, directed by The Research Foundation in memmory of Eva Lee Caldwell

In menopausal women, one of the critical risk factors for breast cancer is obesity/adiposity. It is evident from various studies that leptin, a 16 kDa protein hormone overproduced in obese people, plays the critical role in neovascularization and tumorigenesis in breast and other organs. However, the mechanisms by which obesity influences the breast carcinogenesis remained unclear. In this study, by analyzing different estrogen receptor-α (ER-α)-positive and ER-α-negative BC cell lines, we defined the role of CCN5 in the leptin-mediated regulation of growth and invasive capacity.

We analyzed the effect of leptin on cell viability of ER-α-positive MCF-7 and ZR-75-1 cell lines and ER-α-negative MDA-MB-231 cell line. Additionally, we also determined the effect of leptin on the epithelial-mesenchymal transition (EMT) bio-markers, in vitro invasion and sphere-formation of MCF-7 and ZR-75-1 cell lines. To understand the mechanism, we determined the impact of leptin on CCN5 expression and the functional role of CCN5 in these cells by the treatment of human recombinant CCN5 protein(hrCCN5). Moreover, we also determined the role of JAK-STAT and AKT in the regulation of leptin-induced suppression of CCN5 in BC cells.

Present studies demonstrate that leptin can induce cell viability, EMT, sphere-forming ability and migration of MCF-7 and ZR-75-1 cell lines. Furthermore, these studies found that leptin suppresses the expression of CCN5 at the transcriptional level. Although the CCN5 suppression has no impact on the constitutive proliferation of MCF-7 and ZR-75-1 cells, it is critical for leptin-induced viability and necessary for EMT, induction of in vitro migration and sphere formation, as the hrCCN5 treatment significantly inhibits the leptin-induced viability, EMT, migration and sphere-forming ability of these cells. Mechanistically, CCN5-suppression by leptin is mediated via activating JAK/AKT/STAT-signaling pathways.

These studies suggest that CCN5 serves as a gatekeeper for leptin-dependent growth and progression of luminal-type (ER-positive) BC cells. Leptin may thus need to destroy the CCN5-barrier to promote BC growth and progression via activating JAK/AKT/STAT signaling. Therefore, these observations suggest a therapeutic potency of CCN5 by restoration or treatment in obese-related luminal-type BC growth and progression.

• Breast cancer
• CCN5
• Invasion and migration
• Leptin
• Proliferation

• CCN1
• CCN5
• In situ hybridization
• Probe preparation
• RNA isolation
• Reverse-transcriptase polymerase chain reaction

• Breast cancers
• Differentially expressed genes
• Estrogen receptor positive
• Human epithelial growth factor receptor positive
• Transcriptome analysis

• African-American
• Breast cancer
• European American
• Racial disparity
• Tumor microenvironment
• microRNAs

CCN5/WISP-2 is an anti-invasive molecule and prevents breast cancer (BC) progression. However, it is not well understood how CCN5 prevents invasive phenotypes of BC cells. CCN5 protein expression is detected in estrogen receptor-α (ER-α) -positive normal breast epithelial cells as well as BC cells, which are weakly invasive and rarely metastasize depending on the functional status of ER-α. A unique molecular relation between CCN5 and ER-α has been established as the components of the same signaling pathway that coordinate some essential signals associated with the proliferation as well as delaying the disease progression from a non-invasive to invasive phenotypes. Given the importance of this connection, we determined the role of CCN5 in regulation of ER-α in different cellular settings and their functional relationship. In a genetically engineered mouse model, induced expression of CCN5 in the mammary ductal epithelial cells by doxycycline promotes ER-α expression. Similarly, CCN5 regulates ER-α expression and activity in normal and neoplastic breast cells, as documented in various in vitro settings such as mouse mammary gland culture, human mammary epithelial cell and different BC cell cultures in the presence or absence of human recombinant CCN5 (hrCCN5) protein. Mechanistically, at least in the BC cells, CCN5 is sufficient to induce ER-α expression at the transcription level via interacting with integrins-α6β1 and suppressing Akt followed by activation of FOXO3a. Moreover, in vitro and in vivo functional assays indicate that CCN5 treatment promotes response to tamoxifen in triple-negative BC (TNBC) cells possibly via restoring ER-α. Collectively, these studies implicates that the combination treatments of CCN5 (via activation of CCN5 or hrCCN5 treatment) and tamoxifen as potential therapies for TNBC.

• Wnt signal
• Wnt信号
• cyclin D1
• insulin-like growth factor-1
• matricellular protein
• proliferation
• 细胞周期蛋白D1
• 细胞基质蛋白
• 细胞增殖
• 胰岛素样生长因子甲

• Animals
• CCN Intercellular Signaling Proteins/genetics
• CCN Intercellular Signaling Proteins/metabolism
• Cell Line, Tumor
• Cell Proliferation/genetics
• Cell Survival/genetics
• Humans
• Insulin-Like Growth Factor I/pharmacology
• Islets of Langerhans/drug effects
• Islets of Langerhans/metabolism
• Mice
• Repressor Proteins/genetics
• Repressor Proteins/metabolism
• Wnt Signaling Pathway/drug effects

Breast cancer progression and relapse is conceivably due to tumor initiating cells (TICs)/cancer stem cells. EMT (epithelial-mesenchymal-transition)-signaling regulates TICs’ turnover. However, the mechanisms associated with this episode are unclear. We show that, in triple-negative-breast cancer (TNBC) cells enriched with TICs, CCN5 significantly blocks cellular growth via apoptosis, reversing EMT-signaling and impairing mammosphere formation, thereby blocking the tumor-forming ability and invasive capacity of these cells. To corroborate these findings, we isolated tumor-initiating side populations (SP) and non-side population (NSP or main population) from MCF-7 cell line, and evaluated the impact of CCN5 on these subpopulations. CCN5 was overexpressed in the NSP but downregulated in the SP. Characteristically, NSP cells are ER-α positive and epithelial type with little tumorigenic potency, while SP cells are very similar to triple-negative ones that do not express ER-α- and Her-2 and are highly tumorigenic in xenograft models. The overexpression of CCN5 in SP results in EMT reversion, ER-α upregulation and delays in tumor growth in xenograft models. We reasoned that CCN5 distinguishes SP and NSP and could reprogram SP to NSP transition, thereby delaying tumor growth in the xenograft model. Collectively, we reveal how CCN5-signaling underlies the driving force to prevent TNBC growth and progression.

• Eutheria
• Marsupialia
• cell type evolution
• endometrial stromal cells
• transcriptome evolution

• Breast cancer
• CCN
• CCN6
• CTGF
• CYR61
• Differentiation
• Matricellular
• Metastasis
• NOV
• Stem cells
• WISP1
• WISP2
• WISP3

• CCN
• Degradomics
• ICAT
• Inactive catalytic domain capture
• MMP
• Proteolysis
• Substrate trap
• Yeast two-hybrid

• Antineoplastic Agents, Hormonal/therapeutic use
• Antineoplastic Agents, Hormonal/toxicity
• Breast Neoplasms/drug therapy
• Breast Neoplasms/mortality
• Breast Neoplasms/pathology
• Cell Line, Tumor
• Cell Proliferation/drug effects
• Down-Regulation/drug effects
• Estradiol/toxicity
• Ethanol/pharmacology
• Female
• Humans
• MCF-7 Cells
• Mitogen-Activated Protein Kinase 1/metabolism
• Mitogen-Activated Protein Kinase 3/metabolism
• Neoplasm Proteins/genetics
• Neoplasm Proteins/metabolism
• Proto-Oncogene Mas
• Proto-Oncogene Proteins B-raf/antagonists & inhibitors
• Proto-Oncogene Proteins B-raf/genetics
• Proto-Oncogene Proteins B-raf/metabolism
• RNA, Small Interfering/metabolism
• Receptors, Estrogen/metabolism
• Signal Transduction/drug effects
• Tamoxifen/therapeutic use
• Tamoxifen/toxicity
• Trefoil Factor-1
• Tumor Suppressor Proteins/genetics
• Tumor Suppressor Proteins/metabolism
• Up-Regulation/drug effects

The CCN family of proteins comprises the members CCN1, CCN2, CCN3, CCN4, CCN5 and CCN6. They share four evolutionarily conserved functional domains, and usually interact with various cytokines to elicit different biological functions including cell proliferation, adhesion, invasion, migration, embryonic development, angiogenesis, wound healing, fibrosis and inflammation through a variety of signalling pathways. In the past two decades, emerging functions for the CCN proteins (CCNs) have been identified in various types of cancer. Perturbed expression of CCNs has been observed in a variety of malignancies. The aberrant expression of certain CCNs is associated with disease progression and poor prognosis. Insight into the detailed mechanisms involved in CCN-mediated regulation may be useful in understanding their roles and functions in tumorigenesis and cancer metastasis. In this review, we briefly introduced the functions of CCNs, especially in cancer.

“CCN” is an acronym referring to the first letter of each of the first three members of this original group of mammalian functionally and phylogenetically distinct extracellular matrix (ECM) proteins [i.e., cysteine-rich 61 (CYR61), connective tissue growth factor (CTGF), and nephroblastoma-overexpressed (NOV)]. Although “CCN” genes are unlikely to have arisen from a common ancestral gene, their encoded proteins share multimodular structures in which most cysteine residues are strictly conserved in their positions within several structural motifs. The CCN genes can be subdivided into members developmentally indispensable for embryonic viability (e.g., CCN1, 2 and 5), each assuming unique tissue-specific functions, and members not essential for embryonic development (e.g., CCN3, 4 and 6), probably due to a balance of functional redundancy and specialization during evolution. The temporo-spatial regulation of the CCN genes and the structural information contained within the sequences of their encoded proteins reflect diversity in their context and tissue-specific functions. Genetic association studies and experimental anomalies, replicated in various animal models, have shown that altered CCN gene structure or expression is associated with “injury” stimuli—whether mechanical (e.g., trauma, shear stress) or chemical (e.g., ischemia, hyperglycemia, hyperlipidemia, inflammation). Consequently, increased organ-specific susceptibility to structural damages ensues. These data underscore the critical functions of CCN proteins in the dynamics of tissue repair and regeneration and in the compensatory responses preceding organ failure. A better understanding of the regulation and mode of action of each CCN member will be useful in developing specific gain- or loss-of-function strategies for therapeutic purposes.

Wnt-1 inducible signaling pathway protein-2 (WISP-2) is a member of the CCN family, which is critical for the control of cell morphology, motion, adhesion and other processes involved in tumorigenesis. The expression pattern and clinical significance of WISP-2 in astrocytomas remains unclear. In this study, reverse transcription-polymerase chain reaction was performed to systematically investigate the expression of WISP-2 in 47 astrocytoma tissues of different pathological grades and 10 normal brain tissues. The mRNA expression levels of WISP-2 in the astrocytoma tissues were observed to be significantly higher than those in the normal brain tissues. Furthermore, the upregulation of WISP-2 was found to be associated with astrocytomas of higher pathological grades. Subsequently, 154 astrocytoma and 15 normal brain tissues were analyzed using immunohistochemistry and similar results were obtained. Univariate and multivariate survival analyses were used to determine the correlations between WISP-2 expression and overall survival (OS) and progression-free survival (PFS). The results indicated that the expression of WISP-2 was found to negatively correlate with patient PFS and OS. These results demonstrated that the WISP-2 protein is involved in the pathogenesis and progression of human astrocytomas and may serve as a malignant biomarker of this disease.

• Wnt-1 inducible signaling pathway protein-2
• astrocytoma
• overall survival
• progression-free survival

• 5-aza-2′-deoxycytidine
• GAS1
• cisplatin
• lung cancer
• methylation
• trichostatin A

• Antimetabolites, Antineoplastic/pharmacology
• Antineoplastic Agents/pharmacology
• Apoptosis
• Azacitidine/analogs & derivatives
• Azacitidine/pharmacology
• Carcinoma, Non-Small-Cell Lung/genetics
• Carcinoma, Non-Small-Cell Lung/pathology
• Cell Cycle Checkpoints
• Cell Cycle Proteins/metabolism
• Cell Line, Tumor
• Cell Proliferation
• Cisplatin/pharmacology
• DNA Methylation
• Decitabine
• Drug Resistance, Neoplasm/drug effects
• Epigenesis, Genetic
• GPI-Linked Proteins/metabolism
• Gene Expression Profiling/methods
• Histone Deacetylase Inhibitors/pharmacology
• Humans
• Hydroxamic Acids/pharmacology
• Lung Neoplasms/genetics
• Lung Neoplasms/pathology
• RNA, Messenger/metabolism

The complex signaling networks between cancer cells and adjacent endothelial cells make it challenging to unravel how cancer cells send extracellular messages to promote aberrant vascularization or tumor angiogenesis. Here, in vitro and in vivo models show that pancreatic cancer cell generated unique microenvironments can underlie endothelial cell migration and tumor angiogenesis. Mechanistically, we find that pancreatic cancer cell secreted CCN1/Cyr61 matricellular protein rewires the microenvironment to promote endothelial cell migration and tumor angiogenesis. This event can be overcome by Sonic Hedgehog (SHh) antibody treatment. Collectively, these studies identify a novel CCN1 signaling program in pancreatic cancer cells which activates SHh through autocrine-paracrine circuits to promote endothelial cell migration and tumor angiogenesis and suggests that CCN1 signaling of pancreatic cancer cells is vital for the regulation of tumor angiogenesis. Thus CCN1 signaling could be an ideal target for tumor vascular disruption in pancreatic cancer.

Hypoxia and the hypoxia-inducible factor (HIF) signaling pathway trigger the expression of several genes involved in cancer progression and resistance to therapy. Transcriptionally active HIF-1 and HIF-2 regulate overlapping sets of target genes, and only few HIF-2 specific target genes are known so far. Here we investigated oxygen-regulated expression of Wnt-1 induced signaling protein 2 (WISP-2), which has been reported to attenuate the progression of breast cancer. WISP-2 was hypoxically induced in low-invasive luminal-like breast cancer cell lines at both the messenger RNA and protein levels, mainly in a HIF-2α-dependent manner. HIF-2-driven regulation of the WISP2 promoter in breast cancer cells is almost entirely mediated by two phylogenetically and only partially conserved functional hypoxia response elements located in a microsatellite region upstream of the transcriptional start site. High WISP-2 tumor levels were associated with increased HIF-2α, decreased tumor macrophage density, and a better prognosis. Silencing WISP-2 increased anchorage-independent colony formation and recovery from scratches in confluent cell layers of normally low-invasive MCF-7 cancer cells. Interestingly, these changes in cancer cell aggressiveness could be phenocopied by HIF-2α silencing, suggesting that direct HIF-2-mediated transcriptional induction of WISP-2 gene expression might at least partially explain the association of high HIF-2α tumor levels with prolonged overall survival of patients with breast cancer.

• invasion
• metastasis
• motility
• oxygen
• transcriptional regulation
• tumor

Gastric cancer (GC) is the second leading cause of cancer-related death. The poor survival rate may reflect the relatively aggressive tumor biology of GC. Recently, the importance of the tumor microenvironment in carcinogenesis has emerged. In the tumor microenvironment, tumor cells and the surrounding stromal cells aberrantly secrete matricellular proteins capable of modulating carcinogenesis and regulating metastasis. The Cyr61/CTGF/Nov (CCN) proteins are a family of matricellular proteins with variable roles in many physiological and pathological processes. The evidence suggests that CCN family proteins contribute to GC carcinogenic processes. Here, we briefly review recent research on the effects of CCN family proteins in GC carcinogenesis and the development of new targeted agents in this field.

• Cyr61/CTGF/Nov proteins
• Cysteine-rich angiogenic inducer 61
• Connective tissue growth factor
• Nephroblastoma over-expressed
• Gastric cancer
• Gastric carcinogenesis

• Antibodies, Monoclonal/chemistry
• CCN Intercellular Signaling Proteins/metabolism
• Carcinogenesis
• Connective Tissue Growth Factor/metabolism
• Cysteine-Rich Protein 61/metabolism
• Gene Expression Regulation, Neoplastic
• Humans
• Neoplasm Metastasis
• Nephroblastoma Overexpressed Protein/metabolism
• Protein Sorting Signals
• Protein Structure, Tertiary
• RNA Interference
• Signal Transduction
• Stomach Neoplasms/metabolism
• Stomach Neoplasms/mortality

It has been proposed that the epithelial-mesenchymal transition (EMT) in mammary epithelial cells and breast cancer cells generates stem cell features. WISP2 (Wnt-1-induced signaling protein-2) plays an important role in maintenance of the differentiated phenotype of estrogen receptor-positive breast cancer cells and loss of WISP2 is associated with EMT. We now report that loss of WISP2 in MCF7 breast cancer cells can also promote the emergence of a cancer stem-like cell phenotype characterized by high expression of CD44, increased aldehyde dehydrogenase activity and mammosphere formation. Higher levels of the stem cell markers Nanog and Oct3/4 were observed in those mammospheres. In addition we show that low-cell inoculums are capable of tumor formation in the mammary fat pad of immunodeficient mice. Gene expression analysis show an enrichment of markers linked to stem cell function such as SOX9 and IGFBP7 which is linked to TGF-β inducible, SMAD3-dependent transcription. Taken together, our data demonstrate that WISP2 loss promotes both EMT and the stem-like cell phenotype.

• Animals
• Biomarkers, Tumor/genetics
• Biomarkers, Tumor/metabolism
• Breast Neoplasms/drug therapy
• Breast Neoplasms/metabolism
• Breast Neoplasms/pathology
• CCN Intercellular Signaling Proteins/genetics
• CCN Intercellular Signaling Proteins/metabolism
• Cells, Cultured
• Epithelial-Mesenchymal Transition
• Estrogens/pharmacology
• Female
• Gene Expression Regulation, Neoplastic
• HEK293 Cells
• Humans
• Hyaluronan Receptors/genetics
• Hyaluronan Receptors/metabolism
• Mice
• Mice, Nude
• Neoplastic Stem Cells/drug effects
• Neoplastic Stem Cells/metabolism
• Neoplastic Stem Cells/pathology
• Phenotype
• Repressor Proteins/genetics
• Repressor Proteins/metabolism
• Signal Transduction
• Transforming Growth Factor beta/genetics
• Transforming Growth Factor beta/metabolism

• Apoptosis/genetics
• Biomarkers, Tumor/genetics
• Cell Cycle/genetics
• Cholesterol/blood
• Double-Blind Method
• Estrogens/blood
• Humans
• Isoflavones/therapeutic use
• Male
• Middle Aged
• Phytotherapy
• Placebos
• Prostate-Specific Antigen/blood
• Prostatic Neoplasms/blood
• Prostatic Neoplasms/drug therapy
• Prostatic Neoplasms/genetics
• Prostatic Neoplasms/pathology
• Glycine max/chemistry
• Testosterone/blood
• Time Factors

Canonical and non-canonical Wnt pathways are involved in the genesis of multiple tumors; however, their role in pituitary tumorigenesis is mostly unknown.

This study evaluated gene and protein expression of Wnt pathways in pituitary tumors and whether these expression correlate to clinical outcome.

Genes of the Wnt canonical pathway: activating ligands (WNT11, WNT4, WNT5A), binding inhibitors (DKK3, sFRP1), β-catenin (CTNNB1), β-catenin degradation complex (APC, AXIN1, GSK3β), inhibitor of β-catenin degradation complex (AKT1), sequester of β-catenin (CDH1), pathway effectors (TCF7, MAPK8, NFAT5), pathway mediators (DVL-1, DVL-2, DVL-3, PRICKLE, VANGL1), target genes (MYB, MYC, WISP2, SPRY1, TP53, CCND1); calcium dependent pathway (PLCB1, CAMK2A, PRKCA, CHP); and planar cell polarity pathway (PTK7, DAAM1, RHOA) were evaluated by QPCR, in 19 GH-, 18 ACTH-secreting, 21 non-secreting (NS) pituitary tumors, and 5 normal pituitaries. Also, the main effectors of canonical (β-catenin), planar cell polarity (JNK), and calcium dependent (NFAT5) Wnt pathways were evaluated by immunohistochemistry.

There are no differences in gene expression of canonical and non-canonical Wnt pathways between all studied subtypes of pituitary tumors and normal pituitaries, except for WISP2, which was over-expressed in ACTH-secreting tumors compared to normal pituitaries (4.8x; p = 0.02), NS pituitary tumors (7.7x; p = 0.004) and GH-secreting tumors (5.0x; p = 0.05). β-catenin, NFAT5 and JNK proteins showed no expression in normal pituitaries and in any of the pituitary tumor subtypes. Furthermore, no association of the studied gene or protein expression was observed with tumor size, recurrence, and progressive disease. The hierarchical clustering showed a regular pattern of genes of the canonical and non-canonical Wnt pathways randomly distributed throughout the dendrogram.

Our data reinforce previous reports suggesting no activation of canonical Wnt pathway in pituitary tumorigenesis. Moreover, we describe, for the first time, evidence that non-canonical Wnt pathways are also not mis-expressed in the pituitary tumors.

• Adenocarcinoma/chemistry
• Adenocarcinoma/mortality
• Adenocarcinoma/pathology
• Adult
• Aged
• Aged, 80 and over
• CCN Intercellular Signaling Proteins/analysis
• CCN Intercellular Signaling Proteins/physiology
• Carcinoma, Adenosquamous/chemistry
• Carcinoma, Adenosquamous/mortality
• Carcinoma, Adenosquamous/pathology
• Carcinoma, Squamous Cell/chemistry
• Carcinoma, Squamous Cell/mortality
• Carcinoma, Squamous Cell/pathology
• Carrier Proteins/analysis
• Carrier Proteins/physiology
• Female
• Gallbladder Neoplasms/chemistry
• Gallbladder Neoplasms/mortality
• Gallbladder Neoplasms/pathology
• Humans
• Immunohistochemistry
• Intercellular Signaling Peptides and Proteins/analysis
• Intercellular Signaling Peptides and Proteins/physiology
• Male
• Middle Aged
• Neoplasm Staging
• Proportional Hazards Models
• Repressor Proteins/analysis
• Repressor Proteins/physiology

Most gain of function mutations of tyrosine kinase receptors in human tumours are hemizygous. Gastrointestinal stromal tumours (GIST) with homozygous mutations have a worse prognosis. We aimed to identify genes differentially regulated by hemizygous and heterozygous KIT mutations.

Expression of 94 genes and 384 miRNA was analysed with low density arrays in five NIH3T3 cell lines expressing the full-length human KIT cDNA wild-type (WT), hemizygous KIT mutation with del557-558 (D6) or del564-581 (D54) and heterozygous WT/D6 or WT/D54. Expression of 5 of these genes and 384 miRNA was then analysed in GISTs samples.

Unsupervised and supervised hierarchical clustering of the mRNA and miRNA profiles showed that heterozygous mutants clustered with KIT WT expressing cells while hemizygous mutants were distinct. Among hemizygous cells, D6 and D54 expressing cells clustered separately. Most deregulated genes have been reported as potentially implicated in cancer and severals, as ANXA8 and FBN1, are highlighted by both, mRNA and miRNA analyses. MiRNA and mRNA analyses in GISTs samples confirmed that their expressions varied according to the mutation of the alleles. Interestingly, RGS16, a membrane protein of the regulator of G protein family, correlate with the subcellular localization of KIT mutants and might be responsible for regulation of the PI3K/AKT signalling pathway.

Patterns of mRNA and miRNA expression in cells and tumours depend on heterozygous/hemizygous status of KIT mutations, and deletion/presence of TYR568 & TYR570 residues. Thus each mutation of KIT may drive specific oncogenic pathways.