过表达AnnexinA2抑制食管癌Eca109细胞增殖和迁移

Cell proliferation and migration in Eca109 line suppressed by overexpression of AnnexinA2

目的探讨AnnexinA2的表达对人食管癌细胞Eca109增殖、迁移、细胞周期和凋亡的影响。方法采用基因过表达技术上调Eca109细胞中AnnexinA2的表达,qRT-PCR和Western blot技术检测AnnexinA2 mRNA和蛋白水平变化;赛唑蓝(MTT)比色法研究其对Eca109细胞的存活、增殖能力的影响;采用划痕实验观察其对细胞迁移能力的影响;流式细胞术检测其对Eca109细胞周期及细胞凋亡的影响。结果 qRT-PCR和Western blot检测提示Eca109细胞转染AnnexinA2过表达质粒后mRNA和蛋白质表达水平均显著增高;MTT实验表明AnnexinA2过表达组细胞的增殖能力明显低于对照组(P<0.05),同时细胞迁移能力也显著下降(P<0.05);AnnexinA2在Eca109细胞中高表达将细胞阻滞于G2/M期,对细胞凋亡没有影响。结论 AnnexinA2在食管癌细胞中呈低表达,上调AnnexinA2蛋白水平可以明显抑制食管癌细胞的增殖、迁移能力。

Objective To investigate effects of overexpression of AnnexinA2 on the proliferation, migration and cell cycle and apoptosis in ESCC line Ecal09. Methods Ecal09 cells were transfected with full length ofAnnexinA2 eukaryotic ex- pression vector pCMV-XL5-AnnexinA2. The expression of AnnexinA2 was detected by qRT-PCR and Western blot at mRNA and protein level, respectively, proliferation was examined by MTT assay, migration capacity was evaluated by wound healing assay. In addition, the cell apoptosis and cell-cycle were analyzed by flow cytometry. Results In the group transfected with pCMV-XL5-AnnexinA2, AnnexinA2 expression was significantly increased compared with group transfect- ed with blank vector and normal group. Moreover, there was significant suppression in the cell proliferation and migration ca- pability of Eca109 transfected with pCMV-XL5-AnnexinA2. In addition, up-regulating AnnexinA2 partly caused Eca 109 cell cycle arrest at the G2 phase. Conclusions The present findings demonstrated that up-regulation of AnnexinA2 played an important role in suppressing proliferation and migration of Eca109 cells.