Inhibitory Biological Effects On Hepatoma Cell Hep3B Of SiRNA On Expression P62

Objective: To investigate the biological effects on hepatoma cell Hep3B of p62 by use of p62 siRNA.Methods: The siRNAs were transfected into hepatoma cell Hep3B with Lipofectamine 2000.First, the best siRNA sequence which silent p62 was selected from three siRNA sequences specifically targeting p62 (named as Si-h-GTF2H1₀01, Si-h-GTF2H1₀02 and Si-h-GTF2H1₀03). Then,the exprement were classified four groups: 1,blank control group;2,siRNA group;3,Ncontrol(NC)group;4,Lipofectamine(Lip)group.The expression of p62,XPD,p53,cyclinD1 were detected by RT-PCR and Western blot.The cell cycle were examined by MTT.Results: The best siRNA sequence which silent p62 was Si-h- GTF2H1₀02. Contrast with blank control group , NC group and Lip group ,the expression of p62 mRNA was decreased significantly in siRNA group(p<0.01);the expression of XPD mRNA and p53 mRNA were decreased obviously;but the expression of cyclinD1 mRNA was enhanced obviously.The trends of protein was consistent with mRNA.MTT results showed that cell proliferation increased in siRNA group.Conclusion: p62 gene may initeract with XPD,p53 and cyclinD1 to inhibit cancer cell,and possibly regulate cell proliferation through DNA damage checkpoint.