基础医学与临床 ›› 2014, Vol. 34 ›› Issue (2): 235-239.

• 技术与方法 • 上一篇    下一篇

反转录病毒载体介导的P27kip1促HepG2细胞凋亡

潘永平1,龚辉2,邱梦标2,樊晓慧2,毛红霞2,熊水印2,粟庆娟2   

  1. 1. 南昌大学第四附属医院
    2. 南昌大学 第四附属医院
  • 收稿日期:2013-04-26 修回日期:2013-07-22 出版日期:2014-02-05 发布日期:2014-01-13
  • 通讯作者: 潘永平 E-mail:pypingnc@163.com
  • 基金资助:
    江西省科技厅社会发展支撑项目

Retro-virus mediated human p27kip1 over expression in HepG2 induced cell apoptosis

  • Received:2013-04-26 Revised:2013-07-22 Online:2014-02-05 Published:2014-01-13

摘要: 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株,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株,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。

关键词: 逆转录病毒, P27kip1, HepG2, 表达

Abstract: Objective To study the effect of retro-virus mediated human P27kip1 over expression in HepG2 cells. Method pLNCX vector containing human p27kip1 gene was constructed, and transfected into PA317 package cells, retrovirus containing P27kip1 was collected and infected HepG2 cells, over express P27kip1 cells was selected and the effect on HepG2 was determined. Result high efficient retrovirus vector was constructed, the P27kip1 gene was transferred into HepG2 cells and over expressed in HepG2 cells, and cells over express P72kip1 were caped in G1 phase, and cell growth was inhibited. Conclusion retrovirus vector pLNCX-P27 can efficiently import P27kip1 gene into HepG2 cells, over express P27kip1 inhibited HepG2 cell growth and induce apoptosis.

Key words: retro-virus, P27kip1, HepG2, expression