Des-gamma-carboxy prothrombin increases the expression of angiogenic factors in human hepatocellular carcinoma cells.
OBJECTIVE
Des-gamma-carboxyl prothrombin (DCP) is a serum protein produced by hepatocellular carcinoma (HCC) cells. The aim of this study was to evaluate the angiogenic activity of DCP in HCC cells.


METHODS
The proliferation of HCC cells was measured by 3-[4, 5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay. The growth of HCC cells was also evaluated in vivo by using the xenografts in nude mice. The enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of angiogenic factors in supernatant of cell culture. The expression of angiogenic factors was examined by Western blot analysis and immunohistochemical staining.


RESULTS
DCP displayed the stimulation of HCC cell growth in a dose (5-80 ng/ml) and time (24-96 h) dependent manner. The increase of cell growth was also observed in nude mice bearing well-established, palpable HepG2 and SMMC-7721 xenografts after 2 weeks administration of DCP. HCC cell growth was accompanied by the elevated levels of angiogenic factors. The levels of vascular endothelial growth factor (VEGF), transforming growth factor-alpha (TGF-alpha) and basic fibroblast growth factor (bFGF) in supernatant of SMMC-7721 cells were increased from 47, 126, and 60 pg/10(6) cells/24 h to 400, 208, and 298 pg/10(6) cells/24 h, respectively, after 72 h incubation with 80 ng/ml of DCP. The results of Western blot analysis and immunohistochemical staining of HCC xenografts also showed the significant increase of VEGF, TGF-alpha, and bFGF in HCC cells.


CONCLUSIONS
These results provide the information that DCP is a type of growth factor in progression of HCC.
Gao
Fu-Jun
FJ
Department of Pharmacology, School of Pharmaceutical Sciences, Shandong University, Jinan, Shandong, 250012, China.
Cui
Shu-Xiang
SX
Chen
Ming-Hui
MH
Cheng
Yan-Na
YN
Sun
Li-Rui
LR
Ward
S G
SG
Kokudo
Norihiro
N
Tang
Wei
W
Qu
Xian-Jun
XJ
D042501:Angiogenic Proteins / Q000096:biosynthesis*; D000818:Animals; D015415:Biomarkers; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007150:Immunohistochemistry; D008114:Liver Neoplasms, Experimental / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D011498:Protein Precursors / Q000494:pharmacology* / Q000502:physiology; D011516:Prothrombin / Q000494:pharmacology* / Q000502:physiology
D042501:Angiogenic Proteins; D015415:Biomarkers; D011498:Protein Precursors; C008208:acarboxyprothrombin; D011516:Prothrombin