论文部分内容阅读
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。
Objective: To investigate the regulation of LSD1 by PI3K/AKT/m TOR signaling pathway in esophageal squamous cell carcinoma Eca109 cells. METHODS: LY294002 (0.5, 1.0, 5.0, 10.0, 20.0, and 50.0 μmol/L), RAD001 (0.05, 0.50, 5.00, 10.00, 20.00, and 50.00 μmol/L) were used as inhibitors of different concentrations of PI3K/Akt/m TOR signaling pathways. Eca109 cells were treated for 24 and 48 h respectively, and CCK-8 assay was used to detect cell proliferation; Eca109 cells were then treated with 0, 10, 20 μmol/L LY294002 or RAD001 for 24 h or 20 μmol/L LY294002 or RAD001 for different time (0~). 72 h) Western blot was used to detect changes in PI3K/Akt/m TOR signaling pathway factors, LSD1, and histone H3K4me2 expression. RESULTS: After treated with LY294002 and RAD001, the proliferation of esophageal squamous carcinoma cells was inhibited, and the inhibitory effect was increased with the increase of the concentration (P<0.05). LY294002 inhibited the expression of p-p70S6K in Eca109 cells and upregulated Raptor, Rictor, Expression of p-Akt (Ser473) and histone H3K4me2; RAD001 inhibited the expression of Raptor, Rictor, and p-p70S6K, increased the expression of p-Akt (Ser473), and down-regulated the expression of LSD1 and up-regulated the expression of histone H3K4me2 (P< 0.05). Conclusion: The PI3K/AKT/m TOR signaling pathway regulates LSD1.