下调AEG-1对卵巢癌细胞生物学行为影响的研究

Effect of down-regulation of AEG-1 on biological behavior of ovarian cancer cells

目的探究下调星形细胞上调基因1(astrocyte elevated gene-1,AEG-1)对卵巢癌细胞增殖、凋亡、周期分布及侵袭、迁移能力的影响。方法将卵巢癌细胞分为空白组、沉默组、过表达组,并通过细胞转染建立稳定转染的沉默组、过表达组。MTT检测细胞增殖能力,使用流式细胞仪检测细胞凋亡、周期分布情况,Transwell小室检测细胞侵袭,细胞划痕实验检测细胞迁移,Western blot法检测Bcl-2、PI3K、Akt表达。结果在第24h、48h、72h,沉默组细胞增殖率均低于空白组,凋亡率高于空白组,且过表达组细胞增殖率均高于空白组、沉默组,凋亡率均低于空白组、沉默组(P<0.05)。沉默组细胞处于G1期的比例高于空白组,且过表达组细胞处于G1期的比例均低于空白组、沉默组(P<0.05)。沉默组细胞侵袭细胞数、迁移细胞数均低于空白组(P<0.05);过表达组细胞侵袭细胞数、迁移细胞数均高于空白组、沉默组(P<0.05)。沉默组细胞凋亡相关蛋白Bcl-2、PI3K、Akt相对表达量均低于空白组(P<0.05);过表达组细胞凋亡相关蛋白Bcl-2、PI3K、Akt相对表达量均高于空白组、沉默组(P<0.05)。结论下调AEG-1的表达,能够调控Bcl-2、PI3K、Akt蛋白的表达,抑制卵巢癌细胞增殖,促进卵巢癌细胞凋亡,阻滞细胞周期分布,还能够抑制卵巢癌细胞的侵袭、迁移能力。

Objective To investigate the effects of down-regulated astrocyte elevated gene-1(AEG-1)on the proliferation,apoptosis,cycle distribution,invasion and migration of ovarian cancer cells.Methods Ovarian cancer cells were divided into blank group,silent group and overexpression group,and stable silent group and overexpression group were established by cell transfection.MTT was used to detect cell proliferation;flow cytometry was used to detect cell apoptosis and cycle distribution;cell invasion was detected by Transwell;cell migration was detected by scratch test;Western blot was used to detect the expression of Bcl-2,PI3 K and Akt.Results At 24,48 and 72 hours,the cell proliferation rate of silent group was lower than that of blank group,and the apoptotic rate was higher than that of blank group.The cell proliferation rate of overexpressed group was higher than that of blank group and silent group,and the apoptotic rate was lower than that of blank group and silent group(P<0.05).The proportion of cells in G1 phase in silence group was higher than that in blank group,and the proportion of cells in G1 phase in overexpression group was lower than that in blank group and silence group(P<0.05).The number of invasive cells and migrating cells in silence group was lower than that in blank group(P<0.05),while the number of invasive cells and migrating cells in overexpression group was higher than that in blank group and silence group(P<0.05).The relative expressions of Bcl-2,PI3 K and Akt in silence group were lower than those in blank group(P<0.05),while the relative expressions of Bcl-2,PI3 K and Akt in overexpression group were higher than those in blank group and silence group(P<0.05).Conclusions Downregulation of AEG-1 expression can regulate the expression of Bcl-2,PI3 K and Akt protein,inhibit the proliferation of ovarian cancer cells,promote the apoptosis of ovarian cancer cells,block cell cycle distribution,and inhibit the invasion and migration of ovarian cancer cells.