基础医学与临床 ›› 2015, Vol. 35 ›› Issue (7): 894-899.

• 研究论文 • 上一篇    下一篇

ATF4重组慢病毒抑制C2C12细胞增殖并促凋亡

夏飞1,伍志盟2,李美玲2,邓莉2,胡勤2,郭风劲3   

  1. 1. 重庆医科大学
    2. 重庆医科大学 基础医学院 细胞生物学及遗传学教研室
    3. 重庆医科大学细胞生物学及遗传学教研室
  • 收稿日期:2015-01-15 修回日期:2015-04-27 出版日期:2015-07-05 发布日期:2015-06-23
  • 通讯作者: 郭风劲 E-mail:912446689@qq.com; guofengjin919@vip.sohu.com
  • 基金资助:
    衰老调控因子SIRT6与NF-κB对话在人参皂苷Rg1延缓造血干/祖细胞衰老中的作用;衰老调控因子SIRT6与NF-κB对话在人参皂苷Rg1延缓造血干/祖细胞衰老中的作用;教育部新世纪优秀人才支持计划

ATF4 lentivirus represses proliferation and promotes apoptosis in C2C12 cells

  • Received:2015-01-15 Revised:2015-04-27 Online:2015-07-05 Published:2015-06-23

摘要: 目的 构建人活性转录因子4(ATF4)慢病毒,探讨C2C12细胞成骨分化过程中ATF4基因慢病毒修饰对其增殖和凋亡的影响。方法 构建ATF4重组慢病毒载体质粒,然后与2个包装质粒共转入293T细胞中包装成ATF4慢病毒(LV-ATF4);用病毒感染C2C12细胞,并用流式细胞仪(FCM)检测BMP2诱导C2C12细胞分化时对其增殖凋亡的影响,免疫印迹法检测凋亡相关蛋白的表达,电子显微镜观察凋亡细胞的形态学结构变化。结果 成功构建和包装了ATF4重组慢病毒。FCM检测结果表明,BMP2+LV-ATF4处理组S期细胞(14.89%)低于BMP2+LV-GFP组(30.64%)(P<0.05);BMP2+LV-ATF4处理组细胞凋亡率(31.06%)高于BMP2+LV-GFP组(11.39%)(P<0.05);凋亡相关蛋白的表达与FCM结果一致。 结论 在BMP2诱导C2C12细胞成骨分化时,LV-ATF4慢病毒可促进C2C12细胞的凋亡,抑制其增殖。

关键词: ATF4, 慢病毒, C2C12, 分化, 细胞增殖凋亡

Abstract: Objective To study the effect of lentivirus modification of human activating transcriptional factor4(ATF4) gene on cell apoptosis and cycle in C2C12 cells. Methods The recombinant ATF4 lentivirus were produced in 293T cells following to co-transfection between pWPT-GFP-ATF4 and the lentivirus packaging plasmids. The effect of recombinant lentivirus ATF4 on proliferation and apoptosis in C2C12 cells was detected by flow cytometry(FCM). Western blotting analysis was used to examine the expression of apoptosis-ralated proteins. Results The recombinant lentivirus ATF4 was successfully constructed. FCM analysis showed that under BMP2-induced differentiation condion,the ratio of S phase C2C12 cells in the BMP2+LV-ATF4 group(14.89%) was significantly lower than those in BMP2+LV-GFP group(30.64%)(P<0.05);The apoptosis rate in the BMP2+LV-ATF4 group(31.06%) was significantly higher than those in the BMP2+LV-GFP group(11.39%)(P<0.05).Western blotting analysis showed that the expression of Cleaved Caspase-3,Chop and p-JNK was consistent with results of FCM. Conclusion Under BMP2-induced differentiation condition, LV-ATF4 can suppress the cell proliferation and promote the cell apoptosis in C2C12 cells.

Key words: ATF4, lentivirus, osteogenic differentiation, cell proliferation, cell apoptosis

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