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Amado V, González-Rubio S, Zamora J, Alejandre R, Espejo-Cruz ML, Linares C, Sánchez-Frías M, García-Jurado G, Montero JL, Ciria R, Rodríguez-Perálvarez M, Ferrín G, De la Mata M. Clearance of Circulating Tumor Cells in Patients with Hepatocellular Carcinoma Undergoing Surgical Resection or Liver Transplantation. Cancers (Basel) 2021; 13:2476. [PMID: 34069569 PMCID: PMC8160727 DOI: 10.3390/cancers13102476] [Citation(s) in RCA: 22] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/16/2021] [Revised: 05/14/2021] [Accepted: 05/17/2021] [Indexed: 12/12/2022] Open
Abstract
BACKGROUND In patients with hepatocellular carcinoma (HCC), a complete clearance of circulating tumor cells (CTCs) early after liver transplantation (LT) or surgical resection (LR) could prevent tumor recurrence. METHODS prospective pilot study including patients with HCC who underwent LR or LT from September 2017 to May 2020. Enumeration of CTCs was performed in peripheral blood samples (7 mL) using the Isoflux® system (Fluxion Biosciences) immediately before surgery, at post-operative day 5 and at day 30. A clinically relevant number of CTCs was defined as >30 CTCs/sample. RESULTS 41 HCC patients were included (mean age 58.7 ± 6.3; 82.9% male). LR was performed in 10 patients (24.4%) and 31 patients (75.6%) underwent LT. The main etiology of liver disease was chronic hepatitis C (31.7%). Patients undergoing LR and LT were similar in terms of preoperative CTC count (p = 0.99), but clearance of CTCs within the first month was more pronounced in the LT group. Clusters of CTCs at baseline were associated with incomplete clearance of CTCs at day 30 (54.2% vs. 11.8%, p = 0.005), which in turn impacted negatively on survival (p = 0.038). CONCLUSION Incomplete clearance of CTCs after surgery could be a surrogate marker of HCC aggressiveness.
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Affiliation(s)
- Víctor Amado
- Department of Hepatology and Liver Transplantation, Reina Sofía University Hospital, 14004 Córdoba, Spain; (V.A.); (J.Z.); (R.A.); (J.L.M.); (M.D.l.M.)
- Maimónides Institute of Biomedical Research (IMIBIC), University of Córdoba, 14004 Córdoba, Spain; (S.G.-R.); (M.L.E.-C.); (C.L.); (G.G.-J.); (R.C.); (G.F.)
| | - Sandra González-Rubio
- Maimónides Institute of Biomedical Research (IMIBIC), University of Córdoba, 14004 Córdoba, Spain; (S.G.-R.); (M.L.E.-C.); (C.L.); (G.G.-J.); (R.C.); (G.F.)
- Centro de Investigación Biomédica en Red de Enfermedades hepáticas y digestivas (CIBERehd), 28029 Madrid, Spain
| | - Javier Zamora
- Department of Hepatology and Liver Transplantation, Reina Sofía University Hospital, 14004 Córdoba, Spain; (V.A.); (J.Z.); (R.A.); (J.L.M.); (M.D.l.M.)
- Maimónides Institute of Biomedical Research (IMIBIC), University of Córdoba, 14004 Córdoba, Spain; (S.G.-R.); (M.L.E.-C.); (C.L.); (G.G.-J.); (R.C.); (G.F.)
| | - Rafael Alejandre
- Department of Hepatology and Liver Transplantation, Reina Sofía University Hospital, 14004 Córdoba, Spain; (V.A.); (J.Z.); (R.A.); (J.L.M.); (M.D.l.M.)
- Maimónides Institute of Biomedical Research (IMIBIC), University of Córdoba, 14004 Córdoba, Spain; (S.G.-R.); (M.L.E.-C.); (C.L.); (G.G.-J.); (R.C.); (G.F.)
| | - María Lola Espejo-Cruz
- Maimónides Institute of Biomedical Research (IMIBIC), University of Córdoba, 14004 Córdoba, Spain; (S.G.-R.); (M.L.E.-C.); (C.L.); (G.G.-J.); (R.C.); (G.F.)
| | - Clara Linares
- Maimónides Institute of Biomedical Research (IMIBIC), University of Córdoba, 14004 Córdoba, Spain; (S.G.-R.); (M.L.E.-C.); (C.L.); (G.G.-J.); (R.C.); (G.F.)
- Centro de Investigación Biomédica en Red de Enfermedades hepáticas y digestivas (CIBERehd), 28029 Madrid, Spain
| | | | - Gema García-Jurado
- Maimónides Institute of Biomedical Research (IMIBIC), University of Córdoba, 14004 Córdoba, Spain; (S.G.-R.); (M.L.E.-C.); (C.L.); (G.G.-J.); (R.C.); (G.F.)
| | - José Luis Montero
- Department of Hepatology and Liver Transplantation, Reina Sofía University Hospital, 14004 Córdoba, Spain; (V.A.); (J.Z.); (R.A.); (J.L.M.); (M.D.l.M.)
- Maimónides Institute of Biomedical Research (IMIBIC), University of Córdoba, 14004 Córdoba, Spain; (S.G.-R.); (M.L.E.-C.); (C.L.); (G.G.-J.); (R.C.); (G.F.)
- Centro de Investigación Biomédica en Red de Enfermedades hepáticas y digestivas (CIBERehd), 28029 Madrid, Spain
| | - Rubén Ciria
- Maimónides Institute of Biomedical Research (IMIBIC), University of Córdoba, 14004 Córdoba, Spain; (S.G.-R.); (M.L.E.-C.); (C.L.); (G.G.-J.); (R.C.); (G.F.)
- Department of Hepatobiliary Surgery and Liver Transplantation, Reina Sofía University Hospital, 14004 Córdoba, Spain
| | - Manuel Rodríguez-Perálvarez
- Department of Hepatology and Liver Transplantation, Reina Sofía University Hospital, 14004 Córdoba, Spain; (V.A.); (J.Z.); (R.A.); (J.L.M.); (M.D.l.M.)
- Maimónides Institute of Biomedical Research (IMIBIC), University of Córdoba, 14004 Córdoba, Spain; (S.G.-R.); (M.L.E.-C.); (C.L.); (G.G.-J.); (R.C.); (G.F.)
- Centro de Investigación Biomédica en Red de Enfermedades hepáticas y digestivas (CIBERehd), 28029 Madrid, Spain
| | - Gustavo Ferrín
- Maimónides Institute of Biomedical Research (IMIBIC), University of Córdoba, 14004 Córdoba, Spain; (S.G.-R.); (M.L.E.-C.); (C.L.); (G.G.-J.); (R.C.); (G.F.)
- Centro de Investigación Biomédica en Red de Enfermedades hepáticas y digestivas (CIBERehd), 28029 Madrid, Spain
| | - Manuel De la Mata
- Department of Hepatology and Liver Transplantation, Reina Sofía University Hospital, 14004 Córdoba, Spain; (V.A.); (J.Z.); (R.A.); (J.L.M.); (M.D.l.M.)
- Maimónides Institute of Biomedical Research (IMIBIC), University of Córdoba, 14004 Córdoba, Spain; (S.G.-R.); (M.L.E.-C.); (C.L.); (G.G.-J.); (R.C.); (G.F.)
- Centro de Investigación Biomédica en Red de Enfermedades hepáticas y digestivas (CIBERehd), 28029 Madrid, Spain
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Amado V, Rodríguez-Perálvarez M, Ferrín G, De la Mata M. Selecting patients with hepatocellular carcinoma for liver transplantation: incorporating tumor biology criteria. J Hepatocell Carcinoma 2018; 6:1-10. [PMID: 30613572 PMCID: PMC6306074 DOI: 10.2147/jhc.s174549] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022] Open
Abstract
Liver transplantation (LT) is the optimal therapeutic option for patients with liver cirrhosis and hepatocellular carcinoma (HCC). Due to universal donor shortage, only the patients with limited tumor burden (under the so-called Milan criteria) are considered as potential candidates for LT in most institutions. It is expected that in the near future, more liver grafts will be available for patients with HCC due to the implementation of new direct antivirals against hepatitis C, leaving a prone scenario to consider expanding Milan criteria. A moderate expansion of Milan criteria could be implemented without increasing the risk of tumor recurrence if patients with favorable biological behavior are carefully selected. Incorporating information regarding tumor biology in the decision-making algorithm would result in a more rational use of LT in patients with HCC. In the present review, surrogate markers of tumor biology are critically evaluated as potential tools to be combined with existing radiological criteria. In addition, the current state of liquid biopsy is discussed, as this cutting-edge technology may reshape the management of HCC in the upcoming years.
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Affiliation(s)
- Víctor Amado
- Department of Hepatology and Liver Transplantation, Reina Sofía University Hospital, IMIBIC, CIBERehd, Córdoba, Spain,
| | - Manuel Rodríguez-Perálvarez
- Department of Hepatology and Liver Transplantation, Reina Sofía University Hospital, IMIBIC, CIBERehd, Córdoba, Spain,
| | - Gustavo Ferrín
- Department of Hepatology and Liver Transplantation, Reina Sofía University Hospital, IMIBIC, CIBERehd, Córdoba, Spain,
| | - Manuel De la Mata
- Department of Hepatology and Liver Transplantation, Reina Sofía University Hospital, IMIBIC, CIBERehd, Córdoba, Spain,
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Mann J, Reeves HL, Feldstein AE. Liquid biopsy for liver diseases. Gut 2018; 67:2204-2212. [PMID: 30177542 DOI: 10.1136/gutjnl-2017-315846] [Citation(s) in RCA: 73] [Impact Index Per Article: 10.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/08/2018] [Revised: 07/24/2018] [Accepted: 07/25/2018] [Indexed: 12/12/2022]
Abstract
With the growing number of novel therapeutic approaches for liver diseases, significant research efforts have been devoted to the development of liquid biopsy tools for precision medicine. This can be defined as non-invasive reliable biomarkers that can supplement and eventually replace the invasive liver biopsy for diagnosis, disease stratification and monitoring of response to therapeutic interventions. Similarly, detection of liver cancer at an earlier stage of the disease, potentially susceptible to curative resection, can be critical to improve patient survival. Circulating extracellular vesicles, nucleic acids (DNA and RNA) and tumour cells have emerged as attractive liquid biopsy candidates because they fulfil many of the key characteristics of an ideal biomarker. In this review, we summarise the currently available information regarding these promising and potential transformative tools, as well as the issues still needed to be addressed for adopting various liquid biopsy approaches into clinical practice. These studies may pave the way to the development of a new generation of reliable, mechanism-based disease biomarkers.
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Affiliation(s)
- Jelena Mann
- Newcastle Fibrosis Research Group, Institute of Cellular Medicine, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, UK
| | - Helen L Reeves
- Northern Institute for Cancer Research, Newcastle University, Newcastle upon Tyne, UK
| | - Ariel E Feldstein
- Department of Pediatrics, University of California, San Diego, California, USA
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Simultaneous detection of MCF-7 and HepG2 cells in blood by ICP-MS with gold nanoparticles and quantum dots as elemental tags. Biosens Bioelectron 2016; 90:343-348. [PMID: 27940237 DOI: 10.1016/j.bios.2016.11.030] [Citation(s) in RCA: 59] [Impact Index Per Article: 6.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/24/2016] [Revised: 11/09/2016] [Accepted: 11/11/2016] [Indexed: 01/22/2023]
Abstract
In this work, we demonstrate a novel method based on inductively coupled plasma mass spectrometry (ICP-MS) detection with gold nanoparticles (Au NPs) and quantum dots (QDs) labeling for the simultaneous counting of two circulating tumor cell lines (MCF-7 and HepG2 cells) in human blood. MCF-7 and HepG2 cells were captured by magnetic beads coupled with anti-EpCAM and then specifically labeled by CdSe QDs-anti-ASGPR and Au NPs-anti-MUC1, respectively, which were used as signal probes for ICP-MS measurement. Under the optimal experimental conditions, the limits of detection of 50 MCF-7, 89 HepG2 cells and the linear ranges of 200-40000 MCF-7, 300-30000 HepG2 cells were obtained, and the relative standard deviations for seven replicate detections of 800 MCF-7 and HepG2 cells were 4.6% and 5.7%, respectively. This method has the advantages of high sensitivity, low sample consumption, wide linear range and can be extended to the simultaneous detection of multiple CTC lines in human peripheral blood.
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Abstract
Liver cancer is the fifth most common cancer, but the second leading cause of cancer death, in the world, with more than 700,000 fatalities annually. The major etiology of liver cancer is infection with an hepatotropic virus such as hepatitis B virus or hepatitis C virus infection. While chronic viral infection remains the main cause of liver disease and risk of hepatocellular carcinoma (HCC), rates of nonviral-associated HCC are occurring at an alarmingly increasing rate. Like many cancers, survival rates are closely associated with time of detection. If HCC is caught early, survival rates can be as high as 50%. Regrettably, most cases of HCC are caught late where survival rates can be as low as 2-7%. Thus, there has been great interest in discovering serum biomarkers that could be used to identify those with HCC. To this end, many groups have examined the N-linked glycans to identify changes that occur with HCC. As the liver secretes the vast majority of proteins into the serum, this has often been a starting point for study. In serum, alterations in core fucosylation, outer-arm fucosylation, increased sialylation, and glycan branching have been observed in patients with HCC. Similar findings have been found directly in HCC tissue suggesting that these glycan changes may play a role in tumor formation and development.
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Affiliation(s)
- Anand Mehta
- Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, Pennsylvania, USA
| | - Harmin Herrera
- Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, Pennsylvania, USA
| | - Timothy Block
- Department of Microbiology and Immunology, Drexel University College of Medicine, Doylestown, Pennsylvania, USA
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Li L, Wang HQ, Wang Q, Yang J, Yang JY. Anterior vs conventional approach hepatectomy for large liver cancer: A meta-analysis. World J Gastroenterol 2014; 20:17235-17243. [PMID: 25493040 PMCID: PMC4258596 DOI: 10.3748/wjg.v20.i45.17235] [Citation(s) in RCA: 16] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/04/2014] [Revised: 07/01/2014] [Accepted: 07/25/2014] [Indexed: 02/06/2023] Open
Abstract
AIM: To evaluate the clinical outcomes and safety of anterior- and conventional-approach hepatectomy for patients with large liver tumors.
METHODS: PubMed, EMBASE, Google Scholar and the Cochrane Library databases were searched for randomized controlled trials (RCTs) and controlled clinical trials comparing anterior-approach hepatectomy (AAH) and conventional-approach hepatectomy (CAH). Two observers independently extracted the data using a spreadsheet and assessed the studies for inclusion. Studies that fulfilled the inclusion criteria and addressed the clinical questions of this analysis were further assessed using either fixed effects or random effects models.
RESULTS: Two RCTs and six controlled clinical trials involving 807 patients met the predefined inclusion criteria. A total of 363 patients underwent AAH and 444 underwent CAH. Meta-analysis indicated that the AAH group had fewer requirements for transfusion (OR = 0.37, 95%CI: 0.21-0.63), less recurrence (OR = 0.57, 95%CI: 0.37-0.87), and lower mortality (OR = 0.29, 95%CI: 0.13-0.63). There were no significant differences between AAH and CAH with regard to perioperative complications (OR = 0.94, 95%CI: 0.58-1.51), intraoperative tumor rupture (OR = 0.98, 95%CI: 0.40-2.40), or length of hospital stay (weighted mean difference = -0.17, 95%CI: -2.36-2.02).
CONCLUSION: AAH has advantages of decreased transfusion, mortality and recurrence compared to CAH. It is a safe and effective method for large cancers requiring right hepatectomy.
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Zhang Y, Shi ZL, Yang X, Yin ZF. Targeting of circulating hepatocellular carcinoma cells to prevent postoperative recurrence and metastasis. World J Gastroenterol 2014; 20:142-7. [PMID: 24415867 PMCID: PMC3886003 DOI: 10.3748/wjg.v20.i1.142] [Citation(s) in RCA: 50] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 09/04/2013] [Revised: 10/16/2013] [Accepted: 11/28/2013] [Indexed: 02/06/2023] Open
Abstract
Currently, the main treatment for hepatocellular carcinoma (HCC) involves the surgical removal of tumors or liver transplantation. However, these treatments are often not completely curative, as they are associated with a risk for postoperative recurrence and metastasis. Circulating tumor cells (CTCs) are increasingly recognized as the main source for recurrence and metastasis after radical hepatectomies are performed. Many studies have demonstrated the association between the presence of either pre- or postoperative CTCs and an increased risk for HCC recurrence. To improve the therapeutic outcome of HCC, a personalized, comprehensive and multidisciplinary approach should be considered, involving the application of appropriate diagnostic and therapeutic measures targeting HCC CTCs in different stages throughout the course of treatment. This article proposes some HCC CTC-based strategies for the treatment of HCC, including the monitoring of HCC CTCs before, during and after radical hepatectomy, therapeutic targeting of HCC CTCs, prevention of the generation and colonization of CTCs, as well as the use of CTC indexes for the selection of indications, prediction of prognoses, and planning of individualized therapeutic regimens. Innovation and technological development of therapies targeting CTCs, as well as their translation into clinical practice, will help to effectively reduce postoperative recurrence and metastasis, and significantly prolong the survival of HCC patients.
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MESH Headings
- Animals
- Antineoplastic Agents/adverse effects
- Antineoplastic Agents/therapeutic use
- Biomarkers, Tumor/metabolism
- Carcinoma, Hepatocellular/metabolism
- Carcinoma, Hepatocellular/mortality
- Carcinoma, Hepatocellular/secondary
- Carcinoma, Hepatocellular/surgery
- Chemotherapy, Adjuvant
- Hepatectomy/adverse effects
- Hepatectomy/mortality
- Humans
- Liver Neoplasms/metabolism
- Liver Neoplasms/mortality
- Liver Neoplasms/pathology
- Liver Neoplasms/surgery
- Molecular Targeted Therapy
- Neoplasm Recurrence, Local/prevention & control
- Neoplastic Cells, Circulating/drug effects
- Neoplastic Cells, Circulating/metabolism
- Neoplastic Cells, Circulating/pathology
- Treatment Outcome
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Zhang Y, Li J, Cao L, Xu W, Yin Z. Circulating tumor cells in hepatocellular carcinoma: detection techniques, clinical implications, and future perspectives. Semin Oncol 2012; 39:449-60. [PMID: 22846862 DOI: 10.1053/j.seminoncol.2012.05.012] [Citation(s) in RCA: 63] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
Abstract
Hepatocellular carcinoma (HCC) is a primary liver cancer with a huge challenge in terms of its complex etiology and its management. The fact that the most common site of early tumor recurrence in liver transplantation for HCC is the transplanted allograft strongly suggests that circulating tumor cells (CTCs) are really an active source of HCC metastasis or recurrence. In the past decade, with the tremendous progress in the technology of CTC detection, there is convincing evidence that CTCs have great potential as a marker for metastatic disease and poor prognosis in patients with a malignancy. Currently some interesting and encouraging results have been achieved in HCC CTC detection, although the knowledge about its clinical relevance in HCC is lagging behind other major tumor types. Here we will review existing and developing methodologies for CTC detection, discuss future perspectives, and describe the potential clinical impact of the identification and molecular characterization of CTC subset or circulating cancer stem cells in HCC patients. Particular attention is given to the results based on the HCC CTC study.
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Affiliation(s)
- Yu Zhang
- Molecular Oncology Laboratory, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, China
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Kuo JS, Zhao Y, Schiro PG, Ng L, Lim DSW, Shelby JP, Chiu DT. Deformability considerations in filtration of biological cells. LAB ON A CHIP 2010; 10:837-42. [PMID: 20379567 DOI: 10.1039/b922301k] [Citation(s) in RCA: 79] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/11/2023]
Abstract
Biological cells are highly sensitive to variation in local pressure because cellular membranes are not rigid. Unlike microbeads, cells deform under pressure or even lyse. In isolating or enriching cells by mechanical filtration, pressure-induced lysis is exacerbated when high local fluidic velocity is present or when a filter reaches its intended capacity. Microfabrication offers new possibilities to design fluidic environments to reduce cellular stress during the filtration process. We describe the underlying biophysics of cellular stress and general solutions to scale up filtration processes for biological cells.
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Affiliation(s)
- Jason S Kuo
- Department of Chemistry, University of Washington, Seattle, WA 98195-1700, USA
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Intraoperative blood salvage during liver transplantation in patients with hepatocellular carcinoma: efficiency of leukocyte depletion filters in the removal of tumor cells. Transplantation 2008; 85:863-9. [PMID: 18360269 DOI: 10.1097/tp.0b013e3181671f2e] [Citation(s) in RCA: 55] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023]
Abstract
BACKGROUND Intraoperative blood salvage (IBS) reduces homologous transfusion in orthotopic liver transplantation (OLT), but may carry with it the risk of reinfusing tumor cells in patients with hepatocellular carcinoma (HCC). The use of leukocyte depletion filters (LDFs) for the removal of tumor cells is rarely reported in clinical OLT. The aims of this study were to evaluate the frequency of tumor cell contamination in surgical field during OLT for HCC recipients and to investigate the efficiency of additional LDFs for eliminating tumor cells from IBS. METHODS Thirty-two HCC patients with preoperatively elevated serum alpha-fetoprotein (AFP) underwent OLT. The blood from the surgical field was collected and processed by an autotransfusion device (Cell Saver 5), followed by 2 consecutive LDF filtrations. The HCC cells in IBS samples and filtered samples were determined using a nested RT-PCR technique to detect the AFP mRNA. RESULTS The shed blood samples from 20 (62.5%) of the 32 HCC patients were contaminated with HCC cells and 15 of them remained positive after Cell Saver processing. Patients within the Milan or UCSF criteria were less likely to have HCC cell contamination and the contaminated HCC cells were more likely to be removed by the Cell Saver in these patients as compared to other patients (P<0.01). After filtration through an additional LDF, most cases (13/15) became negative except for those with ruptured tumors (P<0.05). CONCLUSIONS Our results suggest that blood filtration with the LDF can efficiently remove tumor cells and the use of an additional LDF after use of the Cell Saver could markedly reduce the risk of tumor cell reintroduction during the OLT in HCC recipients with nonruptured tumors.
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Albumin mRNA in Plasma Predicts Post-Transplant Recurrence of Patients With Hepatocellular Carcinoma. Transplantation 2008; 85:81-7. [DOI: 10.1097/01.tp.0000298003.88530.11] [Citation(s) in RCA: 30] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/01/2023]
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Tong X, Yang L, Lang JC, Zborowski M, Chalmers JJ. Application of immunomagnetic cell enrichment in combination with RT-PCR for the detection of rare circulating head and neck tumor cells in human peripheral blood. CYTOMETRY PART B-CLINICAL CYTOMETRY 2007; 72:310-23. [PMID: 17205568 DOI: 10.1002/cyto.b.20177] [Citation(s) in RCA: 50] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/30/2023]
Abstract
Detection of rare, circulating tumor cells (CTC's) in human peripheral blood is a potential indicator of prognosis and diagnosis in oncology. Typical methods to detect these CTC's are either by immunocytochemistry (ICCS) or RT-PCR. However without accurate, rapid, and reproducible enrichment processes, these detection techniques are labor intensive and/or unreliable. In this article, a repeatable enrichment process that included a flow-through immunomagnetic cell separation system, the quadrupole magnetic sorter (QMS) was optimized with the aid of a statistical analysis software package. The QMS was operated in a negative mode of operation by immunomagnetically targeting normal human peripheral blood lymphocytes (PBL) through the CD45 surface marker. Three head and neck squamous carcinoma cell lines (HNSCC), Detroit-562, SCC-4, and CAL-27, were used to determine the sensitivity of RT-PCR for the epidermal growth factor receptor (EGFR) in spiked PBL. The detection purity needed for detection was found to be one cell in 10(4), one cell in 10(3), and one cell in 10(5) for the Detroit-562, SCC-4, and CAL-27, respectively. The actual number of cancer cells needed for RT-PCR detection ranged from 30 to 1 cell. To mimic the potential concentration of rare CTC present in peripheral blood of cancer patients, the spiking concentration was chosen to be one cancer cell per 10(5) total leukocytes from healthy donors. Using a single step immunomagnetic labeling, the final, optimized enrichment process produced a 57.6 +/- 30.3-fold (n = 6) enrichment of the rare cancer cells with a final cancer cell recovery of (77.8 +/- 6.6)%.
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Affiliation(s)
- Xiaodong Tong
- Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio 43210, USA
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Cheung ST, Liu CL, Chow JPH, Lee YT, Ip YC, Ho JCY, Fan ST. Preoperative plasma transcript AA454543 level is an independent prognostic factor for hepatocellular carcinoma after partial hepatectomy. Neoplasia 2006; 8:696-701. [PMID: 16984726 PMCID: PMC1584293 DOI: 10.1593/neo.06181] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/30/2023] Open
Abstract
BACKGROUND We have previously reported that tissue expression levels of transcript AA454543 in hepatocellular carcinoma (HCC) are significantly higher than those of normal livers, livers with cirrhosis, and livers with hepatitis. In addition, a higher level of transcript AA454543 in tumor tissues is associated with poor prognosis. We aim to examine whether quantitative measurement of preoperative plasma transcript AA454543 can provide similar prognostic information. PATIENTS AND METHODS Blood samples were obtained from 84 HCC patients before surgery. Real-time quantitative reverse transcription-polymerase chain reaction, using TaqMan system, was employed to measure plasma transcript AA454543 and alpha-fetoprotein (AFP) RNA levels. We assessed their prediction power in prognosis using univariate and multivariate analyses. RESULTS High plasma transcript AA454543 RNA levels were associated with poor overall survival (log-rank test, P < .01). Patients with different plasma AFP RNA levels revealed no difference in overall survival (log-rank test, P = .88). By multivariate Cox regression analysis, plasma transcript AA454543 RNA level (hazard ratio = 4.8, P < .01) and tumor stage (hazard ratio = 1.7, P < .01) were determined to be independent risk factors for the prediction of overall survival. CONCLUSION Preoperative plasma transcript AA454543 RNA level can provide prognostic information for HCC patients receiving curative partial hepatectomy.
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Affiliation(s)
- Siu Tim Cheung
- Center for the Study of Liver Disease, Department of Surgery, The University of Hong Kong, Pokfulam, Hong Kong, China.
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Liu CL, Fan ST, Cheung ST, Lo CM, Ng IO, Wong J. Anterior approach versus conventional approach right hepatic resection for large hepatocellular carcinoma: a prospective randomized controlled study. Ann Surg 2006; 244:194-203. [PMID: 16858181 PMCID: PMC1602174 DOI: 10.1097/01.sla.0000225095.18754.45] [Citation(s) in RCA: 186] [Impact Index Per Article: 9.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/03/2023]
Abstract
OBJECTIVE To evaluate whether major right hepatectomy using the anterior approach technique for large hepatocellular carcinoma (HCC) results in better operative and long-term survival outcomes when compared with the conventional approach technique. SUMMARY BACKGROUND DATA The anterior approach technique has been advocated recently for large right liver tumors. However, its beneficial effects on the operative and survival outcomes of the patients have not been evaluated prospectively. METHODS A prospective randomized controlled study was performed on 120 patients who had large (> or =5 cm) right liver HCC and underwent curative major right hepatic resection during a 57-month period. The patients were randomized to undergo resection of the tumor using the anterior approach technique (AA group, n = 60) or the conventional approach technique (CA group, n = 60). The anterior approach technique involved initial vascular inflow control, completion of parenchymal transection, and complete venous outflow control before the right liver was mobilized. Operative and long-term survival outcomes of the two groups were analyzed. Quantitative assessments of markers of circulating tumor cells at various stages of surgery of the two techniques were also assessed by plasma albumin-mRNA. RESULTS The overall operative blood loss, morbidity, and duration of hospital stay were comparable in both groups. Major operative blood loss of > or =2 L occurred less frequently in the AA group (8.3% vs. 28.3%, P = 0.005). As a result, blood transfusion requirement and number of patients requiring blood transfusion were significantly lower in the AA group. Hospital mortality occurred in 1 patient in the AA group and 6 patients in the CA group (P = 0.114). Median disease-free survival was 15.5 months in the AA group and 13.9 months in the CA group (P = 0.882). Overall survival was significantly better in the AA group (median >68.1 months) than in the CA group (median = 22.6 months, P = 0.006). The survival benefit appeared more obvious in patients with stage II disease and patients with lymphovascular permeation of the tumor. The anterior approach was also found to associate with significantly lower plasma albumin-mRNA levels at various stages of surgery compared with the CA technique. On multivariate analysis, tumor staging, anterior approach hepatic resection, and resection margin involved by the tumor were independent factors affecting overall survival. CONCLUSION The anterior approach results in better operative and survival outcomes compared with the conventional approach. It is the preferred technique for major right hepatic resection for large HCC.
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Affiliation(s)
- Chi Leung Liu
- Centre for the Study of Liver Disease and Department of Surgery, University of Hong Kong, Pokfulam, Hong Kong, China
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15
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Pinzani P, Salvadori B, Simi L, Bianchi S, Distante V, Cataliotti L, Pazzagli M, Orlando C. Isolation by size of epithelial tumor cells in peripheral blood of patients with breast cancer: correlation with real-time reverse transcriptase–polymerase chain reaction results and feasibility of molecular analysis by laser microdissection. Hum Pathol 2006; 37:711-8. [PMID: 16733212 DOI: 10.1016/j.humpath.2006.01.026] [Citation(s) in RCA: 129] [Impact Index Per Article: 6.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 10/31/2005] [Revised: 01/18/2006] [Accepted: 01/20/2006] [Indexed: 11/30/2022]
Abstract
The aim of this study is the counting and the immunomorphological and molecular characterization of circulating tumor cells (CTCs) by the isolation by size of epithelial tumor cells (ISET) method in the peripheral blood of patients with breast cancer. An evaluation of the method's ability to reveal the presence of occult carcinoma cells in blood of a patient with breast cancer was performed and the results compared with those obtained by quantitative real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay for the evaluation of cytokeratin-19 (CK-19) mRNA expression. The feasibility of molecular analysis of CTCs after laser microdissection of filters used in ISET was illustrated, referring to HER-2 amplification. Blood samples drawn from 44 patients with breast cancer were preoperatively analyzed by ISET. From the same samples, total RNA was extracted and submitted to quantitative real-time RT-PCR for the detection of CK-19 mRNA-positive cells using TaqMan technology. HER-2 amplification was measured by real-time RT-PCR on DNA extracted from cells recovered by laser microdissection from 7 selected ISET-positive filters. Of 44 samples, 12 (27%) showed the presence of epithelial cells on the filter (mean +/- SE: 8.5 +/- 2.4 cells per milliliter of blood). A statistically significant agreement (P = .001) was observed between real-time RT-PCR results and those obtained by ISET. With regard to HER-2 amplification, a good correspondence was found between the results obtained from microdissected CTCs and those obtained using DNA extracted from the primary tumor (R = 0.918; P < .01), as well as the immunohistochemistry results. The ISET method allows for the collection of breast carcinoma cells by filtration despite their smaller dimension relative to other carcinoma cell types. The sensitivity and specificity of the method is comparable with those obtained using the quantitative real-time RT-PCR assay for the evaluation of CK-19 mRNA expression. Moreover, the laser microdissection technique allows for the recovery of nucleic acids for further molecular analysis and CTC characterization.
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Affiliation(s)
- Pamela Pinzani
- Clinical Biochemistry Unit, Department of Clinical Physiopathology, University of Florence, 50100 Florence, Italy.
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16
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Wu LQ, Lu Y, Wang XF, Lv ZH, Zhang B, Yang JY. Expression of cancer-testis antigen (CTA) in tumor tissues and peripheral blood of Chinese patients with hepatocellular carcinoma. Life Sci 2006; 79:744-8. [PMID: 16546222 DOI: 10.1016/j.lfs.2006.02.024] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/19/2005] [Revised: 01/23/2006] [Accepted: 02/16/2006] [Indexed: 11/30/2022]
Abstract
To investigate the expression of cancer-testis antigen (CTA) in Chinese patients with hepatocellular carcinoma (HCC), and the relationship between CTA gene expression and clinical indexes, we used one-step reverse transcription polymerase chain reaction (RT-PCR). The expression of the CTA mRNA was investigated in the tissues of HCC and corresponding peripheral blood of 37 patients with HCC. Fifteen samples of cirrhotic tissues and 15 normal tissues were examined with the same method. Two kinds of CTA (SSX-2 and SSX-5) showed high-specific and high-frequent expression in HCC tissues, but neither of them could be detected in adjacent non-HCC tissues. In corresponding peripheral blood of HCC tissues, the positive expression rate of the SSX-2 and SSX-5 mRNA was not very high. No relationship was found between the expression of CTA and clinical indicators such as age, sex, tumor size, TNM staging, serum AFP level and infection with hepatitis virus. In 15 patients with cirrhosis and 15 other non-tumor patients, none of the SSX-2 and SSX-5 mRNA was detected in liver tissue or peripheral blood. High frequency and specificity of CTAs in HCC indicates that their products may be new potential promising targets for antigen-specific immunotherapy of HCC. High frequent co-expression of the two genes in HCC provides a possibility of polyvalent vaccinations for HCC. Specific expression of CTAs was observed in AFP-negative HCC, suggested applying their mRNA as tumor markers to detect circulating HCC cells as adjuvant diagnostic tool and as indicators of recurrence and prognosis.
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MESH Headings
- Adolescent
- Adult
- Aged
- Antigens, Neoplasm/analysis
- Antigens, Neoplasm/blood
- Antigens, Neoplasm/genetics
- Biomarkers, Tumor/blood
- Carcinoma, Hepatocellular/blood
- Carcinoma, Hepatocellular/diagnosis
- Carcinoma, Hepatocellular/genetics
- Case-Control Studies
- Female
- Gene Expression Regulation, Neoplastic
- Humans
- Leukocytes, Mononuclear/chemistry
- Liver Neoplasms/genetics
- Male
- Middle Aged
- Neoplasm Proteins/analysis
- Neoplasm Proteins/blood
- Neoplasm Proteins/genetics
- RNA, Messenger/analysis
- RNA, Messenger/genetics
- Repressor Proteins/analysis
- Repressor Proteins/blood
- Repressor Proteins/genetics
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Affiliation(s)
- Li-Qun Wu
- Department of Hepatobiliary Surgery, Affiliated Hospital of Medical College, Qingdao University, No.16 Jiangsu Rd. Qingdao 266003, China
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17
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Li Y, Dong X, Yin Y, Su Y, Xu Q, Zhang Y, Pang X, Zhang Y, Chen W. BJ-TSA-9, a novel human tumor-specific gene, has potential as a biomarker of lung cancer. Neoplasia 2006; 7:1073-80. [PMID: 16354590 PMCID: PMC1501171 DOI: 10.1593/neo.05406] [Citation(s) in RCA: 65] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/17/2005] [Revised: 08/28/2005] [Accepted: 08/29/2005] [Indexed: 11/18/2022] Open
Abstract
Using bioinformatics, we have identified a novel tumor-specific gene BJ-TSA-9, which has been validated by Northern blot analysis and reverse transcription-polymerase chain reaction (RT-PCR). BJ-TSA-9 mRNA was expressed in 52.5% (21 of 40) of human lung cancer tissues and was especially higher in lung adenocarcinoma (68.8%). To explore the potential application of BJ-TSA-9 for the detection of circulating cancer cells in lung cancer patients, nested RT-PCR was performed. The overall positive detection rate was 34.3% (24 of 70) in peripheral blood mononuclear cells (PBMCs) of patients with various types of lung cancers and was 53.6% (15 of 28) in PBMCs of lung adenocarcinoma patients. In combination with the detection of two known marker genes SCC and LUNX, the detection rate was increased to 81.4%. A follow-up study was performed in 37 patients after surgical removal of tumor mass. Among nine patients with persistent detection of two to three tumor marker transcripts in PBMCs, six patients had recurrence/metastasis. In contrast, 28 patients with transient detection of one tumor marker or without detection of any tumor marker were all in remission. Thus, BJ-TSA-9 may serve as a marker for lung cancer diagnosis and as a marker, in combination with two other tumor markers, for the prediction of the recurrence and prognosis of lung cancer patients.
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MESH Headings
- Antigens, Neoplasm/blood
- Biomarkers, Tumor/blood
- Biomarkers, Tumor/genetics
- Blotting, Northern
- Chromosomes, Human, Pair 8/genetics
- Cloning, Molecular
- Female
- Follow-Up Studies
- Glycoproteins/blood
- Humans
- Lung Neoplasms/blood
- Lung Neoplasms/genetics
- Male
- Neoplasm Proteins/blood
- Neoplasm Proteins/genetics
- Neoplasm Staging
- Neoplastic Cells, Circulating/chemistry
- Phosphoproteins/blood
- Polymerase Chain Reaction
- RNA, Messenger/metabolism
- Sequence Analysis, DNA
- Serpins/blood
- Tumor Cells, Cultured
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Affiliation(s)
- Yunyan Li
- Immunology Department, Peking University Health Science Center, Beijing, China
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18
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Gwak MS, Lee KW, Kim SY, Lee J, Joh JW, Kim SJ, Lee HH, Park JW, Kim GS, Lee SK. Can a leukocyte depletion filter (LDF) reduce the risk of reintroduction of hepatocellular carcinoma cells? Liver Transpl 2005; 11:331-5. [PMID: 15719385 DOI: 10.1002/lt.20346] [Citation(s) in RCA: 30] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
Abstract
During liver transplantation for hepatocellular carcinoma (HCC) patients, HCC could theoretically be introduced into the systemic circulation when salvaged blood is used with an autotransfusion device. Several reports have shown that some types of leukocyte depletion filters (LDFs) could completely reduce the risk for reintroducing some types of tumor cells. In this study, we tested the ability of the LDF (RCEZ1T, Pall Biomedical Co, NY, USA) to reduce the risk for reintroducing HCC cells in vitro by using a very sensitive detection method. We divided the test group into 6 groups: group I was 10 cells, group II was 20 cells, group III was 2 x 10(3) cells, group IV was 2 x 10(5) cells, group V was 2 x 10(6) cells, and group VI was 2 x 10(7) cells. The counted cells in 200 mL saline were passed through the RCEZ1T using the force of gravity. To identify the presence of cells, the pellet was resuspended, and polymerase chain reaction (PCR) was performed. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a housekeeping gene, was used as a primer. In groups I and II, the HCC cells were completely filtered in all experiments. However, in groups III, IV, and V, the HCC cells were not completely filtered in a few of the repeated experiments, with the unfiltered rate of tumor cells being between 8% and 20%. In group VI, the HCC cells were not completely filtered in all the repeated experiments. In conclusion, the RCEZ1T filter markedly reduced the risk for reintroduction of HCC cells. However, at high HCC cell load the filter cannot completely remove all the tumor cells. Further studies are required to assess the impact in clinical settings.
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Affiliation(s)
- Mi Sook Gwak
- Department of Anesthesiology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea
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Zhao L, Mou DC, Leng XS, Peng JR, Wang WX, Huang L, Li S, Zhu JY. Expression of cancer-testis antigens in hepatocellular carcinoma. World J Gastroenterol 2004; 10:2034-8. [PMID: 15237429 PMCID: PMC4572328 DOI: 10.3748/wjg.v10.i14.2034] [Citation(s) in RCA: 25] [Impact Index Per Article: 1.2] [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 cancer-testis (CT) antigens MAGE-1, SSX-1 ,CTp11 and HCA587 genes in hepatocellular carcinoma (HCC) and the possibility of applying these antigens as targets for specific immunotherapy for HCC.
METHODS: Expression levels of MAGE-1, SSX-1, CTp11 and HCA587 mRNA were detected with reverse transcription polymerase chain reaction (RT-PCR) in HCC tissues and corresponding adjacent non-cancerous tissues from 105 HCC patients, 40 samples of cirrhosis and normal liver tissues. Genes of five samples with positive PCR results were sequenced.
RESULTS: Of 105 HCC tissues, MAGE1, SSX-1 ,CTp11 and HCA587 mRNA expressions were detectable in 75.2% (79/105), 72.4% (76/105), 62.9% (66/105) and 56.2% (59/105) of HCC samples, respectively. About 93.3% (98/105), 72.4% (76/105), 48.6% (51/105) and 37.1% (39/105) of HCC tissues positively expressed at least one, two, three, and four members of CT antigens, respectively. Conversely, only SSX-1 could be detectable in 2.9% (3/105) of the corresponding adjacent non-HCC tissues in which no metastatic lesion was found. Of the latter 3 patients, biopsy samples far from tumor were obtained in 2 patients and RT-PCR indicated no expression of SSX-1 mRNA in these two samples. In addition, none of 40 samples of cirrhotic and normal liver tissues expressed CT antigen gene mRNA. DNA sequences confirmed that the RT-PCR products were true target cDNA. No relationship was found between expression of CT antigens and clinico pathological indicators such as age, gender, tumor size, degree of tumor differentiation, serum α-fetoprotein level and infection of hepatitis B virus or hepatitis C virus (P > 0.05).
CONCLUSION: CT antigens genes (MAGE-1, SSX-1, CTp11 and HCA587) are expressed with high percentage and specificity in HCC and their products are promising targets for antigen-specific immunotherapy of HCC. High frequent co-expression of multiple members of CT antigens in HCC provides possibility of polyvalent vaccinations for HCC.
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Affiliation(s)
- Li Zhao
- Center of Hepatobiliary Surgery, People's Hospital, Peking University, 11 XiZhimen Nandajie, West District, Beijing 100044, China
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20
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Vona G, Estepa L, Béroud C, Damotte D, Capron F, Nalpas B, Mineur A, Franco D, Lacour B, Pol S, Bréchot C, Paterlini-Bréchot P. Impact of cytomorphological detection of circulating tumor cells in patients with liver cancer. Hepatology 2004; 39:792-7. [PMID: 14999698 DOI: 10.1002/hep.20091] [Citation(s) in RCA: 160] [Impact Index Per Article: 7.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
Abstract
The clinical impact of circulating tumor cell (CTC) detection is controversial, mainly due to drawbacks of molecular approaches applied to this field. We sought to determine if the specific identification and counting of circulating tumor cells by cytomorphologic analysis has clinical usefulness. Peripheral blood (6 mL), treated using isolation by size of epithelial tumor cells, was obtained from 44 patients with primary liver cancer (PLC) and without metastases, 30 patients with chronic active hepatitis, 39 with liver cirrhosis, and 38 healthy individuals, and followed up for a mean period of 1 year. We searched for beta-catenin mutations in 60 single microdissected CTCs. One patient with liver cancer developed extrahepatic metastases during follow-up. CTCs and microemboli were found in 23 of the 44 patients with liver cancer and in none of the patients with chronic active hepatitis, patients with cirrhosis, or healthy subjects. Their presence was significantly associated with tumor diffusion (P =.0001) and portal tumor thrombosis (P =.006). Both the presence (P =.01) and number (P =.02) of CTCs and microemboli were significantly associated with a shorter survival. Beta-catenin mutations were found in 3 of 60 CTCs, arguing against their impact on the initial step of tumor cell invasion. In conclusion, the highly sensitive and specific detection of CTCs and microemboli may have clinical implications for cancer staging and outcome prediction. We also show the feasibility of molecular studies of individual circulating tumor cells, aimed at identifying gene mutations involved in tumor invasion.
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Affiliation(s)
- Giovanna Vona
- Pasteur/INSERM Unit 370, Faculté de Médecine Necker-Enfants Malades, 156 rue de Vaugirard, 75739 Paris cedex 15, France
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21
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Matsunaga H, Hangai N, Aso Y, Okano K, Kawamura M, Kobayashi K, Kambara H, Hoger JH, Mitsuhashi M. Application of differential display to identify genes for lung cancer detection in peripheral blood. Int J Cancer 2002; 100:592-9. [PMID: 12124810 DOI: 10.1002/ijc.10534] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/10/2022]
Abstract
A blood assay for detection of lung cancer biomarkers could significantly improve cancer patient prognosis and survival rates. Amplified fragment length polymorphism-differential display (AFLP-DD) was used to identify gene transcripts found in lung cancer tissue and the peripheral blood of lung cancer patients. The clones were evaluated for gene expression in lung cancer tissue, peripheral blood of lung cancer patients and healthy volunteers' blood. The isolated gene transcript clones were found to be from the syndecan 1 gene, collagen 1 gene and 2 novel genes. All 4 transcripts were expressed in normal lung tissue, 4 cultured primary lung cells and 6 lung cancer cell lines. RNA was isolated from peripheral blood samples of 69 lung cancer patients. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to test for the presence of cytokeratin 19 and the 4 gene mRNA transcripts in blood RNA. The positive detection rate of at least 1 of the 5 transcripts was 79% for lung adenocarcinoma and 62% for squamous carcinoma. Using RT-PCR, at least 1 of the markers was found in 53% of stage I patients, 100% of stage II, 71% of stage III and 81% of stage IV lung cancer patients. Blood samples from 20 healthy volunteers were also tested, but only 1 of the 5 transcripts was found in 1 patient. These new molecular markers may aid early detection, staging and follow-up of lung cancer patients by RNA isolated from blood.
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22
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Kubota H, Storms RW, Reid LM. Variant forms of alpha-fetoprotein transcripts expressed in human hematopoietic progenitors. Implications for their developmental potential towards endoderm. J Biol Chem 2002; 277:27629-35. [PMID: 12006569 DOI: 10.1074/jbc.m202117200] [Citation(s) in RCA: 34] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022] Open
Abstract
Hematopoietic stem cells have been identified as multipotent cells that give rise to all adult hematopoietic lineages. Although the hematopoietic lineage is derived from the mesodermal germ layer in the embryo, recent data suggest that bone marrow cells with an antigenic profile consistent with that of hematopoietic stem cells can also differentiate to cell types of the endodermal lineages, such as hepatocytes. However, the molecular mechanisms associated with these events are entirely unknown. For decades, alpha-fetoprotein (AFP) has been used as a differentiation marker for endodermal cells, because it was thought that the transcription of AFP mRNA is tightly regulated in a developmental and tissue-specific process. In this report we describe two new variant forms of AFP transcripts in human hematopoietic progenitors that are not expressed in mature cells. The variant AFP (vAFP) cDNA sequences isolated from a multipotent hematopoietic cell line, K562, revealed that the vAFP differed from the authentic transcript, consisting of 15 exons, by replacing exon 1 of AFP with one or two exons located in the 5'-untranslated region of the AFP gene. In addition to the K562 cell line, vAFP transcripts were detected in normal bone marrow, thymus, and brain but were not detected in normal spleen, intestine, liver, or the hepatocellular carcinoma cell line, HepG2. This suggests expression in normal hematopoietic progenitors. This hypothesis was confirmed by the finding that CD34(+)Lin(-) hematopoietic progenitor cells purified from cord blood by flow cytometric sorting also expressed the variant transcripts. These results suggest that some hematopoietic progenitors are in a state that permits them to express certain types of transcripts that have been considered unique to endoderm.
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Affiliation(s)
- Hiroshi Kubota
- Department of Cell and Molecular Physiology and Program in Molecular Biology and Biotechnology, University of North Carolina at Chapel Hill, School of Medicine, Chapel Hill, North Carolina 27599-7038, USA.
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