The Mechanism Of TTF1 Induced Apoptosis In Hepatocellular Carcinoma HepG-2

Objective:Effects observed TTF1 HepG-2 hepatoma cells and its effect on hepatocellular carcinoma HepG-2 cell apoptosis, and further explore its possible mechanism of apoptosis.Methods:MTT colorimetric observation of TTF1 affect the proliferation of hepatoma cells HepG-2 situation, by flow cytometry TTF1 on HepG-2 cell cycle distribution, using immunocytochemistry detection of apoptosis-related gene bc 1-2, bax and p53 protein expression detected by Western blot with Bc 1-2, Bax, P53 protein and Caspase-3, Caspase-8 and Caspase-9 protein expression in the case.Results:TTF1 can inhibit the proliferation of HepG-2 cells, with the time and the concentration of drug increased, the inhibition became more obvious, visible TTF1 on the liver cell line HepG-2 with time and concentration dependent, when concentration was 10μmol/L when the reaction time was 48 h, the inhibition of HepG-2 cells was 47.1%, nearly half of the inhibitory concentration (IC50); TTF1 can change the cell cycle,10μmol/L TTF1 were on the HepG-2 cells for 24 h,48 h,72 h later, G0/G1 phase ratio were 52.0%,59.6%,64.5%, compared with the control group, there were significant differences (P<0.05), indicating that most cells were arrested in G0/G1 phase; 10μmol/ L TTF1 were on the HepG-2 cells for 24 h,48 h,72 h, the DNA content by flow cytometry showed obvious apoptosis peak, the apoptosis rates were 6.3% 7.7%,23.8%, and the apoptosis rate of the control group there was significant difference (P<0.01); compared with the baseline after treatment, apoptosis-related gene bcl-2, bax and p53 protein were significantly change (P<0.01); which Bcl-2, P53 protein was significantly decreased, Bax protein was significantly increased; Caspase-3 and Caspase-9 with the time and the dose increased, zymogen protein band gradually changed thick, cracking products increased gradually, the expression increased gradually, but not Caspase-8 at the same time, over time zymogen protein band became thicker, more pyrolysis products, increased expression, while the change in dose is not obvious.Conclusion:TTF1 can inhibit the proliferation of HepG-2 cells, and possessed some cases-effective, dose-effect relationship and can change cell cycle distribution, and may cause liver HepG-2 cell apoptosis, its mechanism may be related to increases Bax protein expression, reduced Bcl-2, P53 protein expression, and activation of Caspase-9 through the intrinsic apoptotic pathway induced cascade of Caspase-3 induced apoptosis.