外源性APMCF1基因转染对人肝癌细胞系细胞生长和细胞周期的影响

目的：探讨外源性APMCF1基因对人肝癌细胞系（HHCC）细胞生长和细胞周期的影响. 方法： 将人全长APMCF1基因经脂质体包裹转染肝细胞肝癌HHCC细胞,经G418筛选获得转基因癌细胞克隆,采用半定量RT-PCR方法检测APMCF1的表达情况.通过单四唑（MTT）比色,绘制生长曲线,观察APMCF1基因对HHCC细胞生长的影响,流式细胞仪检测 HHCC细胞周期的变化. 结果：半定量RT-PCR结果显示,APMCF1基因被成功地转染至HHCC细胞.稳定表达外源性APMCF1基因的HHCC细胞生长速度明显慢于亲代细胞;细胞周期发生明显改变,G1期细胞比例明显增多,S期细胞比例明显减少. 结论： 外源性APMCF1基因对HHCC细胞生长和细胞周期有明显的抑制作用,可能是一个细胞周期调节基因.

Effects of exogenous APMCF1 gene on the growth and cell cycle of human hepatocarcinoma cell lines cells

Objective: To explore the effects of exogenous APMCF1 gene on the growth and cell cycle of HHCC cells. Methods: A full human APMCF1 gene was cloned into expression vector pcDNA3.0 and transfected into human hepatocellular carcinoma cell HHCC by lipofectamine,and the positive clone was screened by G418.The expression of APMCF1 protein in HHCC cells were examined by semi-quantitative RT-PCR,and its effects on cell growth and cell cycle were observed by MTT and flow cytometer(FCM). Results: Semi-quantitative RT-PCR indicated that APMCF1 had been successfully transfected into HHCC cells.The growth speed of APMCF1 infected cells decreased markedly.Analysis of cell cycle by FCM showed that 85.3% of the infected cells were in G_1 phase and 12.5% in S phase,while 66.4% of the control HHCC cells were in G_1 phase and 24.7% in S phase. Conclusion:Exogenous APMCF1 gene can inhibit the growth and cell cycle of human hepatocellular carcinoma cell HHCC.It may be a cell cycle regulative gene.