The Role Of P53 In Inhibiting The Proliferation Of Hepatic Stellate Cells By Silencing COX-2

ObjectiveObjective to investigate the role of P53 in inhibiting the proliferation of hepatic stellate cells by COX-2-targeting sh RNA.The mechanism of COX-2-targeting shRNA on proliferation of hepatic stellate cells was preliminarily investigated.Methods1.The experiments were divided into negative control group,COX-2 shRNA transfection group,negative control + PFT-a group,COX-2 shRNA transfection+ PFT-a group.2.The COX-2shRNA plasmid was transfected into HSC by liposome LiplofectamineTM2000.The transfection rate was evaluated by fluorescence microscopy.3.MTT was used to detect the proliferation of HSC-T6 cells.4.Q-PCR method was used to detect P53 mRNA and a-SMA(Acta-2)mRNA expression of the results in each group cell.Results1.COX-2shRNA transfection of rat hepatic stellate cells after 48 h,on fluorescence microscope,visible in the cell cytoplasm green fluorescent,transfection rate can reach more than 70%.2.MTT detected the OD490 light absorption value A,which showed that silent COX-2 could inhibit the proliferation of hepatocellular cells.PFT-? can reverse the inhibitory effect of silent COX-2 on the proliferation of hepatic stellate cells.3.Q-PCR results: Transfection group P53 mRNA expression level compared with negative control group,transfection + PFT-a group were raised(P < 0.01);the negative control + PFT-a group compared with negative control group.P53 mRNA expression level was decseased(P < 0.01).,Transfection group of a-SMA mRNA expression level compared with the negative control group,transfection + PFT-a group lowered(P < 0.01);he negative control + PFT-a group compared with negative control group a-SMA mRNA expression level was raised(P < 0.01).ConclusionsCOX-2-targeting shRNA can inhibit the proliferation of HSC-T6,which can be reversed by the P53 signal pathway inhibitor PFT-a,suggesting that COX-2-targeting shRNA may inhibit the proliferation of hepatic stellate cells by affecting the P53 signaling pathway.