抑制Notch和PI3K/Akt信号通路对食管腺癌细胞增殖、侵袭及迁移的影响

目的 探讨Notch和PI3K/Akt两条信号通路对人食管腺癌细胞OE33增殖、侵袭及迁移能力的影响及两条信号通路之间的联系。方法 应用Notch信号通路阻滞剂DAPT和PI3K/Akt信号通路阻滞剂LY294002分别单独和联合处理OE33细胞，设置对照组；Western blot和实时定量PCR法检测OE33细胞中NICD、Hes1、p-Akt、PTEN蛋白和mRNA的表达变化；CCK-8法检测细胞的增殖抑制率；划痕实验用于检测细胞迁移能力的变化；Transwell细胞侵袭实验用于检测细胞的侵袭能力变化。结果 Notch1通路阻滞剂DAPT能减低Notch1通路相关蛋白NICD和Hes1的表达，并能提高p-Akt蛋白的表达；PI3K/Akt路阻滞剂LY294002不仅使p-Akt的蛋白表达水平减低，还能降低Hes1的蛋白表达水平，同时使NICD的蛋白和Hes1的mRNA表达水平增高。DAPT和LY294002联合处理组的NICD、Hes1、p-Akt的蛋白表达均较空白对照组和单药处理组降低，同时细胞的增殖、迁移和转移能力亦明显减弱。各处理组中PTEN的蛋白和mRNA表达水平较对照组均有一定程度的升高。结论 Notch和PI3K/Akt两条信号通路在食管腺癌细胞OE33中可能存在串话, 同时抑制这两种信号通路能有效的抑制OE33的增殖、侵袭及迁移。

Effects on Cell Proliferation,Invasion and Migration of Esophageal Adenocarcinoma by Inhibition of Notch and PI3K /Akt Pathway

Objective To investigate the effects of Notch and PI3K/Akt pathways on cell proliferation, invasion and migration abilities of esophageal adenocarcinoma cell lines OE33 and the association between the two signaling pathways. Methods DAPT as an inhibitor of Notch1 signaling pathway and LY294002 as an inhibitor of PI3K/Akt signaling pathway were given alone and in combination to OE33 cells. Control group were the cells treated without drug. The change of NICD, Hes1, p-Akt and PTEN protein and mRNA expression in OE33 cells were analyzed by Western blot and Real-time Quantitative PCR. The abilities of cell proliferation, migration and invasion were measured respectively by CCK-8, Wound healing assay and Transwell invasion assay. Results DAPT reduced the protein expression levels of NICD and Hes1, while increased the protein expression of p-Akt; LY294002 not only reduced the protein expression of p-Akt but also down-regulated the protein expression of Hes1, meanwhile, up-regulated the level of NICD protein and Hes1 mRNA. Blocking both two pathways resulted in the down-regulation of NICD, Hes1 and p-Akt proteins expression. The combination of two blockers significantly inhibited the abilities of cells proliferation, migration and invasion, compared with the control and single drug treatment group. PTEN protein and mRNA expression levels in each treatment group had increased to some extent. Conclusion The interactions may exist between Notch-1 and PI3K/Akt signaling pathways in OE33 cells, meanwhile, the dual inhibition could effectively weaken the proliferation, invasion and migration of OE33 cells.