反转录病毒载体介导的P27^(kip1)促HepG2细胞凋亡

Retro-virus mediated human P27^(kip1) over expression in HepG2 induced cell apoptosis

目的研究反转录病毒介导的P27kip1基因过表达对HepG2细胞的影响。方法构建携带有人P27kip1基因的反转录病毒载体,经脂质体介导转染PA317包装细胞,G418筛选获得稳定产毒细胞系,病毒感染HepG2细胞,筛选出P27kip1阳性克隆,Western blot检测P27kip1在HepG2中的表达,并通过细胞形态学观察、MTT、FCW等方法,检测P27kip1对HepG2细胞的影响。结果成功构建了含有人P27kip1基因的pLNCX-P27反转录病毒载体并导入包装细胞获得了稳定产毒细胞系,病毒感染HepG2后P27kip1基因可在细胞内高表达,过表达P27kip1基因的细胞生长速度受阻,较多细胞阻滞于G1期,且凋亡增多。结论构建的pLNCX-P27载体能稳定高效地将P27kip1基因导入HepG2细胞,过表达P27kip1可抑制细胞生长,加速凋亡。

Objective To study the effect of retro-virus mediated human P27^kip1 over expression in HepG2 cells. Methods pLNCX vector containing humanP27^kip1 gene was constructed, and transfected into PA317 package cells, retrovirus containing P27^kip1was collected and infected HepG2 cells, P27^kip1 overexpressed cells were selected and the effect on HepG2 was determined. Results High efficient retrovirus vector was constructed, the P27^kip1 gene was transferred into HepG2 cells and over expressed in HepG2 cells. Cells over express P72kipl were caped in G1 phase, and cell growth was inhibited. Conclusions Retrovirus vector pLNCX-P27 can efficiently import P27^kip1gene into HepG2 cells, over expression of P27^kip1 inhibited HepG2 cell growth and induce apoptosis.