The Effect Of PIG11Gene Silencing On The Apoptosis Of Human Hepatocellular Carcinoma HepG2Cells

Objective:Construct the HepG2cell line low expressing PIG11gene and use the HepG2cell line high expressing PIG11gene which was constructed before by our research group, to research on the function of PIG11gene in liver cancer cell and the role of which to promote apoptosis. Explore the mechanism of expression PIG11gene for HepG2cells lead to apoptosis.Methods:Construct miRNA expression plasmid and transform them to the Competent bacteria for DNA sequencing analysis. Transfecting these vectors into HepG2cell by lipofectamineTM2000. The expression of PIG11cells was detected by RT-PCR and PIG11protein was analyzed by Western Blot. Culture five groups cell:HepG2cell> pLXSN-PIG11-HepG2cell、pLXSN-HepG2cell、miR-PIG11-HepG2cell、miR-HepG2cell. Compare apoptosis of five groups cell using Hoechst33342dyeing fluorescence,and the flow cytometer scans (FACS),and Western blot analysis.Results:Chromatin condensation, nuclear fragmentation and apoptotic body were observed in pLXSN-PIG11-HepG2cell group.Flow cytometer scans showed that the apoptosis ratio of HepG2、miR-PIG11-HepG2、miR-HepG2、pLXSN-PIG11-HepG2、 pLXSN-HepG2is5.72%±0.81%、1.34%±0.71%、5.10%±0.40%.34.83%±2.29%、5.34%±0.60%.The apoptosis ratio of HepG2cell with PIG11gene high expression was higher while that of HepG2cell with PIG11gene low expression was the opposite than others.Western blot showed that the expression of caspase8was higher while that of HepG2cell with PIG11gene low expression were lower than others.And the expression of Bcl-2was lower while that of HepG2cell with PIG11gene low expression was higher than others.Conclusions:The apoptosis ratio of HepG2cell with PIG11gene low expression was lower than HepG2cells apoptosis induced by PIG11was related to caspase8and Bcl-2possiblely.