敲除PPP5C对肝癌细胞生长影响的研究

Knock-down of PPP5C impairs cell cycle progression and inhibits cell growth of hepatocellular carcinoma

目的探讨丝氨酸/苏氨酸蛋白磷酸酶5(PPP5C)在肝细胞癌(hepatocellular carcinoma,HCC)进展过程中所发挥的作用。方法采用实时PCR技术分析人类六个不同HCC细胞系中PPP5C的表达水平,使用慢病毒介导的短发夹RNA(shRNA)抑制PPP5C在HCC细胞系Hep G2和Bel-7404中的表达,观察PPP5C在HCC细胞的增殖、克隆的形成及周期的进展方面所起到的作用。结果在慢病毒短发夹PPP5C感染的细胞中,PPP5C的信使RNA水平有显著下降。在PPP5C降低之后,HCC细胞的增殖和克隆形成能力都严重受损。此外,细胞周期分析显示PPP5C降低后,Hep G2细胞在G0/G1阶段和G2/M阶段被抑制。结论 PPP5C能够在HCC细胞中表达,而PPP5C的降低能够抑制HCC细胞的增殖、克隆的形成及周期的进展。RNAi导致的PPP5C下降对于HCC的治疗可能是一种潜在的新型治疗策略。

Objective To explore the role of PPP5C in hepatocellular carcinoma（ HCC） cell growth. Methods Expression levels of PPP5C were determined in six different HCC cell lines by real-time PCR. Lentivirus mediated short hairpin RNA（ shRNA） was employed to silence PPP5C expression in HCC cell lines HepG2 and Bel-7404. And the functional roles of PPP5C in HCC proliferation,clony formation and cell cycle were observed. Results The mRNA levels of PPP5C were down-regulated significantly in Lv-sh PPP5C infected cells. Both the proliferation and colony formation ability of HCC cells were severely impaired after knock-down of PPP5C. Moreover,cell cycle analysis showed PPP5C depletion led to HepG2 cells arrest in G_0/ G_1 phase and G_2/ M phase. Conclusion PPP5C is expressed in HCC and knock-down of PPP5C could inhibit HCC cell proliferation and colony formation as well as cell cycle progression. Down-regulation of PPP5C by RNAi may provide a potential new therapeutic strategy for HCC.