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本研究观察钙蛋白酶抑制剂(ALLN和calpain inhibitorⅣ)对纤连蛋白(FN)诱导的MCF-10A乳腺上皮细胞-间质转化(EMT)的影响。FN作用MCF-10A细胞48 h后,采用划痕修复实验检测MCF-10A细胞的迁移能力;基质胶包被的transwell小室实验检测细胞的侵袭能力;Western blot法检测波形蛋白(vimentin)、E-钙粘着蛋白(E-cadherin)、锌指蛋白(snail)和钙蛋白酶-2(calpain-2)的表达。研究结果显示,FN诱导MCF-10A细胞形态发生改变;显著增加MCF-10A细胞的迁移和侵袭能力;上调calpain-2、vimentin和snail蛋白表达,下调E-cadherin蛋白水平。ALLN和calpain inhibitorⅣ能显著抑制FN诱导的MCF-10A细胞形态变化、迁移和侵袭能力增强、vimentin、snail和calpain-2蛋白表达上调及E-cadherin蛋白表达下调。以上研究结果表明,FN诱导MCF-10A乳腺上皮细胞发生EMT可能与上调calpain-2的表达有关,ALLN和calpain inhibitorⅣ能够抑制FN诱导的上皮间质转化。
This study was to investigate the effect of calpain inhibitors (ALLN and calpain inhibitor Ⅳ) on fibronectin (FN) -induced MCF-10A mammary epithelial-mesenchymal transition (EMT). The migration ability of MCF-10A cells was detected by scratch repair assay after FN treatment of MCF-10A cells for 48 h. The invasion ability of MCF-10A cells was detected by matrigel-coated transwell chamber assay. The expressions of vimentin, E- E-cadherin, snail and calpain-2 expression were measured. The results showed that FN induced morphological changes of MCF-10A cells, significantly increased the migration and invasion ability of MCF-10A cells, up-regulated the expression of calpain-2, vimentin and snail proteins and down-regulated the expression of E-cadherin. ALLN and calpain inhibitor Ⅳ could significantly inhibit the morphological changes of MCF-10A cells induced by FN, and enhanced the ability of migration and invasion, up-regulated the expression of vimentin, snail and calpain-2 and downregulated the expression of E-cadherin. The above results show that FN induced EMT in MCF-10A mammary epithelial cells may be related to the up-regulation of calpain-2 expression, ALLN and calpain inhibitorⅣ can inhibit FN-induced epithelial-mesenchymal transition.