Preliminary Discussion On The Mechanism Of RN181 Affecting The Apoptosis Of The Hepatocellular Carcinoma HepG2 Cells

Objective: To investigate the effects of overexpression of RN181 on the apoptosis and autophagy of hepatocellular carcinoma cell line Hep G2.Methods: By transient transfection, expression regulation in the hepatocellular carcinoma cell line Hep G2, conventional culture transfected 24 h and 48 h after, under a fluorescence microscope observation and photographed, Western blot test RN181 expression level.Overexpression of RN181 successful regulation after flow cytometric detection of apoptosis situation.Western blot was used to detect the apoptosis related protein Bax, bcl-2 and autophagy related protein Beclin1 and p62 expression. Join the p38-MAPK excited agent fennel mildew element(anisomycin), the phosphorylation of p38-MAPK was detected by Western blot flow cytometry was used to detect the apoptosis rate, Western blot was used to detect the apoptosis related protein cleaved Caspase-3, Bax and bcl-2 expression, detection of autophagy related protein Beclin1 and p62 expression.Results: 1. Overexpression of RN181, flow cytometry results showed that overexpression of RN181 group(the apoptosis rate was 54.76 ± 1.8%), and control group(apoptosis rate of 40.9 ± 0.78%), increased significantly(P < 0.01), suggesting that overexpression of RN181 induced apoptosis.Western blot results in Hep G2 hepatoma cells, overexpression of RN181 group compared with the control group, the expression of Bax remarkably increased(P < 0.05), and significantly decrease the expression of Bcl-2(P < 0.05), suggesting that overexpression of RN181 can promote the apoptosis of hepatocellular carcinoma cell line Hep G2. 2. Western blot suggest that overexpression of RN181 group compared with the control group, the expression of Beclin1 remarkably increased(P < 0.05), and significantly decrease the expression of p62(P < 0.01), suggesting that overexpression of RN181 could promote autophagy in hepatocellular carcinoma cell line Hep G2.3. Western blot showed that, compared with the control group, over expression of RN181 significantly decreased the phosphorylation of p-38MAPK(P-p38) expression level protein(P < 0.05), while anisomycin treatment alone(80 ng/m L, 0.5 h), the expression of P-p38 protein was significantly increased(P < 0.05). Addition of anisomycin, Western blot showed that, compared with the over expression RN181 group, expression levels of Bax, Caspase-3 Cleaved and Beclin1 protein were significantly decreased in both RN181 and anisomycin treated groups(P < 0.01), while Bcl-2 protein expression was significantly increased(P < 0.01), and p62 protein expression was significantly increased(P < 0.001);The results above suggest that p-38 MAPK pathway mediates the apoptosis and autophagy of RN181 in Hep G2 cells.Conclusion: RN181 promotes the apoptosis and autophagy of hepatocellular carcinoma Hep G2 by inhibiting the p38-MAPK pathway.