Phenotypic and functional characteristics of endothelial cells derived from human liver cancer.
- Author:
Lian-Qiu WU
1
;
Wen-Jian ZHANG
;
Li-Ya YE
;
Zhi-Hua YANG
;
Jin-Ning LOU
Author Information
- Publication Type:Journal Article
- MeSH: Antigens, CD34; metabolism; Carcinoma, Hepatocellular; genetics; metabolism; pathology; Cell Proliferation; drug effects; Cell Shape; Cells, Cultured; Endothelial Cells; metabolism; pathology; ultrastructure; Gene Expression; Humans; Integrin alphaVbeta3; metabolism; Integrins; metabolism; Intercellular Adhesion Molecule-1; metabolism; Lipoproteins, LDL; metabolism; Liver Neoplasms; genetics; metabolism; pathology; Lung; blood supply; metabolism; pathology; Matrix Metalloproteinase 2; metabolism; Microscopy, Electron, Scanning; Neovascularization, Pathologic; metabolism; pathology; Phenotype; Plasminogen Activator Inhibitor 1; metabolism; Platelet Endothelial Cell Adhesion Molecule-1; metabolism; Receptors, Tumor Necrosis Factor, Type I; metabolism; Receptors, Vitronectin; metabolism; Tissue Plasminogen Activator; metabolism; Tumor Cells, Cultured; Tumor Necrosis Factor Decoy Receptors; metabolism; Tumor Necrosis Factor-alpha; pharmacology; von Willebrand Factor; metabolism
- From: Chinese Journal of Oncology 2007;29(6):419-423
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo analyze the phenotypic and functional characteristics of endothelial (T3A) cells derived from human hepatocellular cell carcinoma.
METHODSEndothelial cells were isolated from human hepatocellular carcinoma specimens. The identification of T3A cells was performed by checking von Willebrand Factor (vWF), CD31, CD34 and Dil-Ac-LDL uptake. The cell surface fenestrations, a specific morphological feature of tumor derived EC, were investigated by scanning and transmission electron microscopy. The phenotypic characteristics of T3A cells were analyzed by fluorescence-activated cell sorter (FACS) and were further conformed by real-time PCR at transcription level. Furthermore, tumor necrosis factor alpha (TNFalpha)-induced cytotoxicity was evaluated by 3-(4, 5-dimethythiazolyl) -2, -diphenyl-2H-tetrazolium-bromide (MTT) assay; Matrix metalloproteinase secretion was detected by zymography; Angiogenic ability in vitro was analyzed by culturing T3A cells in three-dimensional Matrigel plug. Coagulant and fibrinolytic activities were detected by enzyme-linked immunosorbent assay (ELISA).
RESULTSThe isolated T3A cells exhibited classic "spindle-shape" morphology and monolayer growth and contact inhibition properties. Immunofluorescent staining showed that T3A cells expressed vWF, CD31, CD34, and uptake of Dil-Ac-LDL at a high level. The cell surface fenestrations were observed on T3A cells by scanning and transmission electron microscopy. By FACS and real-time PCR, T3A cells were found to express alphav3, alphavbeta5 and TNF receptor p75 at high levels, and TNF receptor p55 and ICAM-1 at low levels, as compared with those in human liver sinusoidal endothelial cells (LSEC). In response to TNFalpha, LSEC exhibited a dose-dependent cytotoxicity, while T3A cells were resistant. Gelatin zymography showed that MMP-2 activity was higher in T3A cells than that in LSEC. In a three-dimensional plug of Matrigel, T3A cells exhibited stronger angiogenic ability as compared with LSEC. In addition, T3A cells released more tissue factor (TF), tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor (PAI-1) and urine plasminogen activator (u-PA) than LSEC in response to TNFalpha.
CONCLUSIONTumor-derived endothelial cells are phenotypically and functionally different from those derived from normal liver tissue.
