Study On The Effects Of Apatinib On The PI3K Singaling Pathway And The Apoptosis Of HepG-2 Hepatocarcinoma Cells

Objective : The aim of this study is to investigate whether Apatinib inhibits the proliferation of hepG-2 hepatoma cells and promotes cells' apoptosis through regulating the PI3 K signaling pathway.Method: Using different concentrations of Apatinib(0,1,2,4,6,8 and 16 ?mol/L)to treat hepG-2 hepatoma cells that grown i n log phase in vitro for 24 hours,the apoptosis rate was measured by flow cytometry,and the relationship between drug concentration and apoptosis was observed.Then using CCK8 kit to detect the ef fect of apatinib on cell proliferation activity,and calculated the dru g IC50(semi-inhibitory concentration).After treated cells with apati nib in IC50 for 0,12,24,48 hours,the apoptosis was measured b y flow cytometry to reveal the relationship between drug action tim e and apoptosis.Then using different concentrations of Apatinib(0,I C50-,IC50 and IC50+)to treat hepG-2 hepatoma cells for 24 hours,the expression levels of Bax,Bcl-2 and PI3 K,p-PI3 k were detectedby Western-blot(WB),and the relative RNA expression levels of Bax and Bcl-2 were detected by real-time PCR(qRT-PCR).Results:1.CCK8 results showed that compared with the control group(0 ?mol/L),the OD values of each concentration group decreased a nd the difference was statistically significant(p<0.01).The higher t he concentration was,the more the OD value decreased.And the I C50 is 13.228(?13)?mol/L calculated by spss20.0.2.Flow cytology results showed that compared with the contro l group(0?mol/L),the apoptotic rate of each group increased,the difference was statistically significant(p<0.01).The higher the conc entration was,the more the apoptotic rate decreased.After treated c ells with Apatinib in IC50(13 ?mol/L after completion)for 0,12,24,and 48 hours,compared with the control group(0 hour),the a poptotic rate of each group increased,the difference was statisticall y significant(p<0.01),and the value increased gradually with time.3.Western Blot results showed that compared with the control group(0?mol/L),the expression of Bax increased while the Bcl-2decreased in each group,the difference was statistically significant(p<0.01).Andcompared with the group(0?mol/L),the protein expr ession of p-PI3 K decreased in each group,the difference was statistically significant(p<0.01),while the expression of PI3 K protein wa s not obvious and non-directional.4.The PCR results showed that compared with the control gro up(0 ?mol/L),the relative RNA expression of Bax in each group increased while the Bcl-2 decreased,the difference was statistically significant(p<0.01).Conclusion:1.Apatinib can promote the apoptosis of hepG-2 hepatoma cell s in vitro.2.Apatinib promote the apoptosis of cells by enhancing the ex pression of the apoptosis gene Bax and inhibiting the expression of the anti-apoptotic gene Bcl-2.3.Apatinib promote the apoptosis of cells by inhibitting the ph osphorylation of PI3 K pathway.