地高辛对缺氧诱导的乳腺癌细胞上皮间质转化和侵袭的影响

Effect of digoxin on hypoxia-induced epithelial-mesenchymal transition and invasion in human breast carcinoma MCF-7 cells

目的:本研究旨在探究地高辛对缺氧诱导乳腺癌细胞上皮间质转化、迁移和侵袭能力的影响,并探讨其分子机制。方法:选取人乳腺癌MCF-7细胞作为研究对象,采用CoCl_2模拟化学缺氧条件,采用细胞划痕实验测量细胞迁移率,采用Transwell侵袭实验检测细胞侵袭力,采用Western blot方法检测人乳腺癌MCF-7细胞缺氧诱导因子-1α(HIF-1α)、Snail、E-cadherin和vimentin蛋白表达的变化。结果:缺氧使MCF-7细胞从多角形上皮形态转变成梭形间质细胞形态,细胞间隙增大,而在地高辛作用下缺氧的MCF-7细胞未发生明显的上皮间质转化。细胞划痕实验和Transwell侵袭实验结果显示,经CoCl_2处理的MCF-7细胞的迁移和侵袭能力均显著增强(P<0.01),地高辛可抑制CoCl_2诱导的细胞迁移和侵袭(P<0.01)。与control组相比,CoCl_2组细胞的HIF-1α、Snail和vimentin蛋白表达水平显著升高(P<0.01),E-cadherin的蛋白表达水平显著降低(P<0.01);CoCl_2+digoxin组细胞的HIF-1α、E-cadherin和vimentin蛋白表达水平与control组相比差异均无统计学显著性,Snail蛋白表达水平虽略高于control组(P<0.05),但与CoCl_2组细胞相比显著降低(P<0.01)。结论:地高辛可通过下调HIF-1α和Snail蛋白表达抑制缺氧诱导的MCF-7细胞上皮间质转化和侵袭。

AIM： To investigate the effect of digoxin on hypoxia-induced epithelial-mesenchymal transition（ EMT）,migration and invasion in human breast carcinoma MCF-7 cells. METHODS： MCF-7 cells were treated in vitro with a chemical hypoxia inducer cobalt chloride（ CoCl_2） to imitate hypoxia. Cell migration was observed by wound healing assay,and cell invasion was measured by Transwell invasion assay. The protein levels of hypoxia-inducible factor-1α（ HIF-1α）,Snail,E-cadherin and vimentin in MCF-7 cells were detected by Western blot. RESULTS： Digoxin inhibited CoCl_2-induced EMT and reversed the mesenchymal phenotype. CoCl_2 enhanced the abilities of migration and invasion（ P〈0. 01）,significantly decreased the expression of E-cadherin and increased the expression of HIF-1α,Snail and vimentin（ P〈0. 01）,but these effects were blocked by digoxin. CONCLUSION： Digoxin inhibits CoCl_2-induced EMT and invasion most likely via HIF1-α-Snail signaling pathway.