Changes in proliferation and apoptosis of human hepatocellular carcinoma cells transfected with Cyclin D1 siRNA

ZHANG Hongying; HU Lilin; YUAN Wenfeng; DING Hao

doi:10.3969/j.issn.1002-266X.2017.35.007

Abstract：Objective To observe the changes of proliferation and apoptosis of human hepatocellular cells transfected with Cyclin D1 siRNA,and to explore the mechanism. Methods The cultured human hepatoma cell line Hep3B was di-vided into the blank control group,negative control group,and experimental group. The negative control group and the ex-perimental group were transfected with NC siRNA and Cyclin D1 siRNA by liposomes,and the cells were harvested after 48 h. MTT was used to detect cell proliferation;the flow cytometry was used to observe apoptotic cells and we calculated apop-tosis rate;the Caspase-3 protein in cells of each group was detected by immunohistochemistry;the mRNA and protein ex-pression levels of hepatitis B virus x protein (HBx),xeroderma pigmentosum D (XPD),Bcl-2,and Bax in cells of each group were detected by RT-PCR and Western blotting,respectively. Results Compared with the blank control group and the negative control group,the cell proliferation ability of the experimental group decreased,the apoptosis rate increased, and the relative expression of Caspase-3 increased (all P < 0. 01). Compared with the blank control group and the negative control group,the mRNA and protein expression levels of HBx and Bcl-2 in the experimental group were down-regulated, while those of XPD and Bax were up-regulated (all P < 0. 01). Conclusion The proliferation of human hepatoma cells transfected with Cyclin D1 siRNA is inhibited and the apoptosis increases,and the mechanism may be related to the down-regulation of HBx and Bcl-2 expression,and up-regulation of XPD and Bax expression.