Expression Of BFGF + MRNA Induced By Angiotensin Ⅱ In Human Hepatocellular Carcinoma Cells

Objective:(1) To investigate the effects of angiotensin II (Ang II) on the expression of basic fibroblast growth factor (bFGF) in human hepatic cancer cell line HepG2.(2) To compare the reliability between CCK-8method and BrdU ELISA method in measure the proliferation of human hepatic cancer cell lines HepG2.Methods:(1) Cultured HepG2were treated by Ang II with various concentrations and collected at different time points within which contained3control groups, introducing Enzyme-Linked immunosorbent assay (ELISA) and real time polymerase chain reaction(PCR) method to detect the expression of bFGF in HepG2cell lines respectively, meantime apply cck-8method to detect the influence of Ang â…¡ to HepG2cells.(2) Perform CCK-8and BrdU-ELISA method, as well as immunofluorescence method to exam the the proliferation of cells Stimulated by using Ang II with concentration of10-7mol/L24hours when the degree of the cell confluence reaches60%and80%.Results:(1) Ang II stimulated the proliferation of HepG2cells by increased bFGF protein expression and enhanced the expression of bFGF mRNA (P<0.01),which was concentration and time dependent.The effect reach its maximum value that the concentration of Ang II reaches10-7mol/L and the time reaches48hours and which could be suppressed by Angiotensin II type1receptor (AT1R) antagonist.(2) The ratio are1.30and1.59Comparing with their own control group respectively by CCK-8and BrdU-ELISA method under the circumstance that the cell confluence reaches60%which have an approximate estimated value; and the ratio are1.02and1.40when the cell confluence reaches80%which the immunofluorescence method reveals that the group with stimulation of Ang II have a obviously higher cell proliferation than control group.Conclusion:(1) The expression of bFGFmRNA can be induced by Ang II in human hepatic cancer cell line HepG2through AT1R, which may play a definite role in tumor growth and metastasis.(2) BrdU-ELISA method is able to present a more objective situation of the cell proliferation compared with CCK-8method.