丙型肝炎病毒核心蛋白对HePG_2细胞增殖和细胞周期的影响

The effect of Hepatitis C virus core protein on morphology, proliferation and cell cycle of HePG_2 cells

目的 观察核心蛋白对细胞增殖和细胞周期调控的影响。方法 观察表达HCV核心蛋白的细胞株HePG2 C和作为对照的细胞株HePG2 CMV的细胞形态 ;绘制细胞生长曲线 ;检测细胞周期 ;用流式细胞仪、免疫组化染色检测PCNA ,P16 ,P2 7,P5 3;半定量RT PCR检测P16 ,P2 7,P5 3mRNA。结果 转染重组质粒PBK HVCC的HePG2 C细胞与转染空载体PBK CMV的HePG2 CMV细胞相比细胞形态未见明显变化。生长曲线显示HePG2 细胞在转染PBK HCVC和空载体PBK CMV后 ,生长速度无明显变化。检测转染空载体的细胞HePG2 CMV 81.3%进入G0 /G1期 ,7.7%进入S期 ;而转染PBK HCVC的细胞HePG2 C 73%进入G0 /G1期 ,13.7%进入S期。在进一步分子机制的探讨中发现HePG2 C细胞PCNA、P5 3表达显著增加 ,P16、P2 7表达显著降低。同样的变化也发生在转录水平。结论 HCV核心蛋白可能通过调节某些基因的转录与表达 ,增加细胞DNA合成 ,扰乱细胞周期 ,进而参与致癌过程。

Objective To explore the effect of Hepatitis C virus core protein on morphology, proliferation and cell cycle of HePG 2 cells. Methods The morphology of HePG 2 C and HePG 2 CMV cells was observed by microscope. The growth curve was also made. Flow cytometry was used to determine cell cycle, the expression and transcription of PCNA, P53, P27, P16 were determined by immunohistochemical method. Results No obvious difference was found between HePG 2 C and HePG 2 CMV cells in morphology and growth rate. Flow cytometry analysis showed that 81.3% HePG 2 CMV cells was in G 0 G 1 phase, 7.7%was in S phase; and 73% HePG 2 C cells was in G 0-G 1 phase, 13.7% in S phase. HCV core protein was found to promote PCNA and P53 expression and transcription, and depress P16 and P27 expression and transcription. Conclusion HCV core protein could promote DNA synthesize, cell cycle, and involves in carcinogenesis.