食管鳞癌Eca109细胞中PI3K/AKT/mTOR信号通路对LSD1的调控作用

Regulation effects of PI3K/AKT/m TOR signaling pathway on LSD1 in Eca109 cells

目的:探讨食管鳞癌Eca109细胞中PI3K/AKT/m TOR信号通路对LSD1的调控作用。方法:用不同浓度的PI3K/Akt/m TOR信号通路抑制剂LY294002(0.5、1.0、5.0、10.0、20.0和50.0μmol/L)、RAD001(0.05、0.50、5.00、10.00、20.00和50.00μmol/L)分别处理Eca109细胞24、48 h,采用CCK-8实验检测细胞增殖;然后分别用0、10、20μmol/L LY294002或RAD001处理Eca109细胞24 h或20μmol/L LY294002或RAD001处理不同时间(0~72 h),采用Western blot检测PI3K/Akt/m TOR信号通路因子、LSD1及组蛋白H3K4me2表达的变化。结果:LY294002、RAD001处理后,食管鳞癌细胞增殖受抑,且随浓度的增加,抑制作用有增强的趋势(P<0.05);LY294002抑制Eca109细胞中p-p70S6K的表达,上调Raptor、Rictor、p-Akt(Ser473)以及组蛋白H3K4me2的表达;而RAD001抑制了Raptor、Rictor、p-p70S6K表达,增加p-Akt(Ser473)的表达,并且下调LSD1表达,上调组蛋白H3K4me2的表达(P<0.05)。结论:PI3K/AKT/m TOR信号通路对LSD1存在调控作用。

Aim： To explore the regulation effects of PI3K/AKT/m TOR signaling pathway on LSD1 in esophageal squamous cell carcinoma（ESCC）Eca109 cells.Methods：Eca109 cells were treated with PI3K/AKT/mT OR signaling pathway inhibitor LY294002 or RAD001 with different concentrations for 24,48 h,then the proliferation of cells was detected by CCK-8 assay.Eca109 cells were treated with LY294002 or RAD001 with different concentration for 24 h,or treated with LY294002 or RAD001 at the same concentration for different time,then the expressions of proteins in PI3K/AKT/mT OR signaling pathway,LSD1 and histone H3K4me2 were detected by Western blot,respectively.Results：The proliferation of Eca109 cells was significantly inhibited by LY294002 or RAD001,and the inhibition effects increased along with the increase of the concentration（P〈0.05）;LY294002 inhibited the expression of p-p70S6K and promoted the expressions of Raptor,Rictor,p-Akt（Ser473）and histone H3K4me2;while RAD001 inhibited the expressions of Raptor,Rictor,p-p70S6K,up-regulated the expressions of p-Akt（Ser473）and histone H3K4me2,and down-regulated that of LSD1（P〈0.05）.Conclusion：The PI3K/AKT/mT OR signaling pathway has regulation effects on LSD1.