Rab25基因下调snail表达抑制人乳腺癌MDA-MB-231细胞上皮-间充质转化能力的研究

Rab25 gene down-regulates snail expression and inhibits epithelial mesenchymal transformation of human breast cancer MDA-MB-231 cells

目的探讨Rab25基因对TGF-β诱导的乳腺癌MDA-MB-231细胞增殖、侵袭及上皮-间充质转化(epithelial-mesenchymal transition,EMT)能力的抑制作用并探索其分子机制。方法将实验分为3组:对照组、TGF-β组和si-Rab25组;构建TGF-β诱导的MDA-MB-231细胞EMT模型;CCK-8实验检测细胞的增殖情况;Transwell小室检测细胞侵袭情况;利用Western blot实验检测各组相关蛋白的变化情况。结果TGF-β刺激MDA-MB-231细胞后,Rab25基因高表达;si-Rab25组细胞迁移能力及侵袭能力减弱(33.49%±2.93%),与TGF-β组(57.48%±3.62%)相比降低41.7%,差异有统计学意义(P<0.05)。si-Rab25组细胞增殖能力(115.32%±5.69%)减弱,与TGF-β组(153.21%±4.17%)相比降低24.73%,差异有统计学意义(P<0.05)。si-Rab25组中MDA-MB-231细EMT相关蛋白表达较TGF-β组差异有统计学意义(P<0.05)。si-Rab25组p-AKT及Snail蛋白的表达与TGF-β组相比显著降低,差异有统计学意义(P<0.05)。结论Rab25基因在MDA-MB-231细胞中处于高表达状态,沉默Rab25基因能在激活AKT信号通路,抑制Snail蛋白的表达,调控EMT相关蛋白表达,抑制其EMT转化。

Objective To probe into Rab25 Gene’s Effect on TGF-βinhibition of proliferation,invasion and epithelial mesenchymal transformation(EMT)of breast cancer MDA-MB-231 cells and explore its molecular mechanism.Methods The experiment was divided into three groups:control group,TGF-βGroup and si-Rab25 group.TGF-βinduced MDA-MB-231 cell model of EMT was built.CCK-8 assay was used to detect cell proliferation.Transwell assay was used to detect the ability of cell invasion and migration.Western blot was used to detect the changes of related proteins in each group.Results After stimulating MDA-MB-231 cells with TGF-β,Rab25 gene was highly expressed.Compared with TGF-βgroup(57.48±%3.62%),the migration ability and invasion ability of cells in si-Rab25 group(33.49%±2.93%)decreased by 41.7%,with a significant difference(P<0.05).Compared with TGF-βgroup(153.21%±4.17%),the proliferation ability of cells in si-Rab25 group(115.32%±5.69%)decreased by 24.73%,with a significant difference(P<0.05).The expression of MDA-MB-231 fine EMT related protein in si-Rab25 group was significantly different from that in TGF-βgroup(P<0.05).The expression of p-AKT and Snail protein in si-Rab25 group was significantly lower than that in TGF-βgroup(P<0.05).Conclusions Rab25 gene is highly expressed in MDA-MB-231 cells.Silencing Rab25 gene can activate AKT signal pathway,inhibit Snail protein expression,regulate EMT related protein expression,and inhibit EMT transformation.