Inhibition of Hela cell proliferation by LKB1 through AMPK signaling pathway

Abstract:

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.