Pirh2对人肝细胞肝癌细胞增殖的影响

Effect of Pirh2 on cell proliferation of hepatocellular carcinoma

目的探讨泛素连接酶Pirh2对人肝细胞肝癌(HCC)细胞增殖的影响。方法流式细胞仪测定血清饥饿释放过程中SMMC-7721和Hep3B细胞的周期分布情况,并用Western blot检测增殖过程中Pirh2及p27Kip1蛋白表达。在SMMC-7721和Hep3B细胞中转染Pirh2shRNA,cellcounting kit-8(CCK-8)和流式细胞分析检测转染后细胞增殖的能力,Western blot检测细胞周期调控因子的表达。结果细胞饥饿72h,两种细胞周期进程停滞在G1/G0期,血清释放后增殖到S期。在此过程中,Pirh2表达上调,p27Kip1表达下调。转染Pirh2shRNA组相较于对照组和转染空载体质粒组,CCK-8和流式细胞分析显示细胞的增殖能力下降及细胞的周期进程明显受到抑制(P<0.05)。结论 Pirh2参与HCC细胞的周期进程调控,有望成为HCC治疗的新靶点。

Objective To invetigate the effect of E3 ligase Pirh2 on the proliferation process of hepatocellular carcinoma（HCC）. Methods The distribution of cell cycle of SMMC-7721 and Hep3B cells during serum starvation and release was examined by flow cytometery. Western blot was used to detect the protein levels of Pirh2 and p27^Kip1 of both ceils. After transfected short hairpin RNA （shRNA） targeting Pirh2, the proliferation and cell cycle distribution of both cells were detected by cell counting Kit-8 （CCK-8） and flow cytometery. The expressions of regulation factor for cell cycle were detected by Western blot Results After serum deprivation for 72 h, both SMMC-7721 and Hep3B cells were arrested in G1/G0 phase, and then serum release stimulated the proliferation to S phase. The expression of Pirh2 was increased during the progress, while the expression of p27^Kip1 was decreased. Moreover, cell proliferation was greatly inhibited and cell cycle was arrested in G0/G1 phase in Pirh2-shRNA transfected SMMC-7721 and Hep3B cells compared with untransfected and control shRNA （P〈0. 05）. Conclusion Pirh2 participates in the regulation and control of cell cycle of human HCC and could become a potential new target for the treatment of HCC.