引用本文:梁小燕,王丕龙,陶小红.LKB1通过AMPK信号通路抑制Hela细胞增殖的机制研究[J].重庆医科大学学报,2013,(1):45~47
LKB1通过AMPK信号通路抑制Hela细胞增殖的机制研究
Inhibition of Hela cell proliferation by LKB1 through AMPK signaling pathway
DOI:
中文关键词:  细胞周期  抑癌基因  LKB1  AMPK
英文关键词:cell cycle  tumor suppressor  LKB1  adenosine monophosphate activated protein kinase
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作者单位
梁小燕,王丕龙,陶小红 重庆医科大学附属第一医院消化内科重庆 400016 
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中文摘要:
      目的:观察LKB1基因对Hela细胞株的抑制作用,探讨其可能的分子机制。方法:体外培养LKB1表达缺失的Hela细胞株,脂质体介导LKB1质粒转染,G418抗性筛选获得稳定表达株。研究细胞周期的变化,Western blot检测磷酸化Rb、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、AMPKα(adenosine monophosphate activated protein kinase α)及磷酸化AMP-Kα的蛋白表达,分析AMPK抑制剂Compound C对PCNA表达的影响。结果:成功构建稳定表达LKB1的Hela细胞系,与空质粒对照组细胞相比,LKB1蛋白稳定表达后使Hela细胞G1期比例明显增加而S期比例减少,Rb蛋白的磷酸化水平、PCNA表达水平明显降低,AMPKα的磷酸化水平明显增强,当用Compound C抑制AMPK活性后,PCNA蛋白表达水平明显提高。结论:在Hela细胞中,LKB1通过激活AMPK信号通路,抑制Rb蛋白的磷酸化,使细胞周期停滞在G1期,抑制细胞增殖。
英文摘要:
      Objective:To observe the inhibitory effect of LKB1 gene on Hela cell proliferation and to investigate the underlying mech-anism. Methods:Hela cell lines lacking LKB1 expression were cultured in vitro and were transfected with liposome mediated LKB1 plasmid. Stable Hela cell lines constitutively expressing LKB1 protein were established by using G418 antibiotic selection pressure. Cell cycle profile and phase distribution were analyzed by flow cytometry. Protein levels of phosphorylated Rb,proliferating cell nu-clear antigen(PCNA),adenosine monophosphate activated protein kinase α(AMPK α) and phosphorylated AMPKα were analyzed by Western blot. AMPK inhibitor Compound C was also used to examine the effect of AMPK inhibition on cell proliferation. Results:Hela cell lines expressing LKB1 protein were successfully established. Constitutive expression of LKB1 in Hela stable cells led to a delayed G1/S transition,accompanied by a decrease in Rb phosphorylation. Reduced expressions of PCNA and elevated phosphoryla-tion of AMPKα were also observed in Hela stable cells. Besides,the expression of PCNA was greatly increased in Hela stable cells treated with AMPK inhibitor Compound C. Conclusion:Expression of LKB1 protein in Hela cells inhibited cell proliferation,which is mediated by LKB1 induced AMPK activation.
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