Human Hepatocellular Carcinoma Tumor–derived Endothelial Cells Manifest Increased Angiogenesis Capability and Drug Resistance Compared with Normal Endothelial Cells

Purpose: Increasing evidence indicates that tumor-derived endothelial cells (TEC) possess a distinct and unique phenotype compared with endothelial cells (NEC) from adjacent normal tissue and may be able to acquire resistance to drugs. The aim of this study was to investigate the angiogenesis activity and response to drug treatment of TECs and NECs derived from human hepatocellular carcinoma (HCC). Experimental Design: TECs or NECs were isolated from HCC or adjacent normal liver tissue using anti-CD105 antibody coupled to magnetic beads. The phenotypic and functional properties of endothelial cells were characterized by testing the expression of CD105, CD31, CD144, vascular endothelial growth factor receptor-1, vascular endothelial growth factor receptor-2, and von Willebrand factor, and the ability of DiI-Ac-LDL-uptake and tube formations. CD105+ TECs were compared with CD105+ NECs and human umbilical vein endothelial cells (HUVEC) by examining their ability to proliferate, motility, ability to adhere to tumor cells, response to tumor conditioned medium, and reactions to the chemotherapy drugs Adriamycin and 5-fluorouracil and the antiangiogenic drug sorafenib. Results: Compared with CD105+ NECs and HUVECs, CD105+ TECs showed increased apoptosis resistance and motility and proangiogenic properties. Meanwhile, CD105+ TECs had a greater ability to adhere to tumor cells and survive in the tumor environment. Moreover, CD105+ TECs acquired more resistance to Adriamycin, 5-fluorouracil, and sorafenib than CD105+ NECs and HUVECs. Conclusions: TECs possessed enhanced angiogenic activity and resistance to chemotherapeutic drugs and an angiogenesis inhibitor, and may provide a better tool for studying tumor angiogenesis and antiangiogenesis drugs in HCC.

[1]  Zhao-You Tang,et al.  Biological characteristics of fluorescent protein-expressing human hepatocellular carcinoma xenograft model in nude mice , 2008, European journal of gastroenterology & hepatology.

[2]  R. Jove,et al.  Sorafenib inhibits signal transducer and activator of transcription 3 signaling associated with growth arrest and apoptosis of medulloblastomas , 2008, Molecular Cancer Therapeutics.

[3]  G. Invernici,et al.  Transforming growth factor-beta1 and CD105 promote the migration of hepatocellular carcinoma-derived endothelium. , 2008, Cancer research.

[4]  J. Bruix,et al.  Molecular targeted therapies in hepatocellular carcinoma , 2008, Hepatology.

[5]  M. Klagsbrun,et al.  Calcification of multipotent prostate tumor endothelium. , 2008, Cancer cell.

[6]  S. Rafii,et al.  Phase II trial evaluating the clinical and biologic effects of bevacizumab in unresectable hepatocellular carcinoma. , 2008, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[7]  L. Ellis,et al.  Endoglin (CD105): A Marker of Tumor Vasculature and Potential Target for Therapy , 2008, Clinical Cancer Research.

[8]  M. Klagsbrun,et al.  Tumor Endothelial Cell Abnormalities , 2008 .

[9]  M. Peakman,et al.  Human pancreatic islet endothelial cells express coxsackievirus and adenovirus receptor and are activated by coxsackie B virus infection , 2007, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[10]  Adrian L Harris,et al.  Anti-Dll4 therapy: can we block tumour growth by increasing angiogenesis? , 2007, Trends in molecular medicine.

[11]  S. Seaman,et al.  Genes that distinguish physiological and pathological angiogenesis. , 2007, Cancer cell.

[12]  Anil K Sood,et al.  Gene alterations identified by expression profiling in tumor-associated endothelial cells from invasive ovarian carcinoma. , 2007, Cancer research.

[13]  V. Pistoia,et al.  Tumor origin of endothelial cells in human neuroblastoma. , 2007, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[14]  J. Folkman.  Angiogenesis: an organizing principle for drug discovery? , 2007, Nature reviews. Drug discovery.

[15]  R. Poon,et al.  Angiogenesis and antiangiogenic therapy in hepatocellular carcinoma. , 2006, Cancer letters.

[16]  D. McDonald,et al.  Rapid vascular regrowth in tumors after reversal of VEGF inhibition. , 2006, The Journal of clinical investigation.

[17]  D. Amadori,et al.  Phase II study of sorafenib in patients with advanced hepatocellular carcinoma. , 2006, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[18]  Ronald A. DePinho,et al.  Hepatocellular carcinoma pathogenesis: from genes to environment , 2006, Nature Reviews Cancer.

[19]  Alona Muzikansky,et al.  Phase II study of gemcitabine and oxaliplatin in combination with bevacizumab in patients with advanced hepatocellular carcinoma. , 2006, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[20]  Lei Xu,et al.  Onset of abnormal blood and lymphatic vessel function and interstitial hypertension in early stages of carcinogenesis. , 2006, Cancer research.

[21]  E. Dejana,et al.  Endothelial cadherins and tumor angiogenesis. , 2006, Experimental cell research.

[22]  J. Gillespie,et al.  Tumor-associated endothelial cells display GSTP1 and RARβ2 promoter methylation in human prostate cancer , 2006, Journal of Translational Medicine.

[23]  Dhara N. Amin,et al.  Tumor endothelial cells express epidermal growth factor receptor (EGFR) but not ErbB3 and are responsive to EGF and to EGFR kinase inhibitors. , 2006, Cancer research.

[24]  A. Sapino,et al.  Isolation and characterization of human breast tumor-derived endothelial cells. , 2006, Oncology reports.

[25]  Wei Wang,et al.  Correlation between CD105 expression and postoperative recurrence and metastasis of hepatocellular carcinoma , 2006, BMC Cancer.

[26]  L. Qin,et al.  Overexpression of Platelet-Derived Growth Factor Receptor α in Endothelial Cells of Hepatocellular Carcinoma Associated with High Metastatic Potential , 2005, Clinical Cancer Research.

[27]  Oriol Casanovas,et al.  Drug resistance by evasion of antiangiogenic targeting of VEGF signaling in late-stage pancreatic islet tumors. , 2005, Cancer cell.

[28]  S. Fan,et al.  Clinicopathological and prognostic implications of endoglin (CD105) expression in hepatocellular carcinoma and its adjacent non-tumorous liver. , 2005, World journal of gastroenterology.

[29]  P. Rieckmann,et al.  Isolation and Culture of Microvascular Endothelial Cells from Gliomas of Different WHO Grades , 2005, Journal of Neuro-Oncology.

[30]  Dhara N. Amin,et al.  Tumor-Associated Endothelial Cells with Cytogenetic Abnormalities , 2004, Cancer Research.

[31]  R. Taub.  Liver regeneration: from myth to mechanism , 2004, Nature Reviews Molecular Cell Biology.

[32]  D. McDonald,et al.  Abnormalities of basement membrane on blood vessels and endothelial sprouts in tumors. , 2003, The American journal of pathology.

[33]  B. Teicher,et al.  Endothelial precursor cells as a model of tumor endothelium: characterization and comparison with mature endothelial cells. , 2003, Cancer research.

[34]  Seth M Steinberg,et al.  A randomized trial of bevacizumab, an anti-vascular endothelial growth factor antibody, for metastatic renal cancer. , 2003, The New England journal of medicine.

[35]  G. Camussi,et al.  Altered angiogenesis and survival in human tumor‐derived endothelial cells , 2003, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[36]  Rakesh K Jain,et al.  Molecular regulation of vessel maturation , 2003, Nature Medicine.

[37]  P. Choyke,et al.  Imaging of angiogenesis: from microscope to clinic , 2003, Nature Medicine.

[38]  Shant Kumar,et al.  CD105 is important for angiogenesis: evidence and potential applications , 2003, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[39]  Kenneth J. Hillan,et al.  Angiogenesis-Independent Endothelial Protection of Liver: Role of VEGFR-1 , 2003, Science.

[40]  Rakesh K Jain,et al.  Abnormalities in pericytes on blood vessels and endothelial sprouts in tumors. , 2002, The American journal of pathology.

[41]  H. Wada,et al.  Evaluation of angiogenesis in non-small cell lung cancer: comparison between anti-CD34 antibody and anti-CD105 antibody. , 2001, Clinical cancer research : an official journal of the American Association for Cancer Research.

[42]  S. Lemeshow,et al.  Genetic model of multi-step breast carcinogenesis involving the epithelium and stroma: clues to tumour-microenvironment interactions. , 2001, Human molecular genetics.

[43]  K. Kinzler,et al.  Genes expressed in human tumor endothelium. , 2000, Science.

[44]  M. Hendrix,et al.  Molecular biology of breast cancer metastasis Molecular expression of vascular markers by aggressive breast cancer cells , 2000, Breast Cancer Research.

[45]  R K Jain,et al.  Openings between defective endothelial cells explain tumor vessel leakiness. , 2000, The American journal of pathology.

[46]  P. Carmeliet.  Mechanisms of angiogenesis and arteriogenesis , 2000, Nature Medicine.

[47]  A. Giatromanolaki,et al.  Intratumoral angiogenesis: a new prognostic indicator for stage I endometrial adenocarcinomas? , 1999, Oncology research.

[48]  J. Folkman.  Angiogenesis in cancer, vascular, rheumatoid and other disease , 1995, Nature Medicine.

[49]  S. Kumar,et al.  Breast carcinoma: comparative study of tumor vasculature using two endothelial cell markers. , 1994, Journal of the National Cancer Institute.