整合素αvβ3抑制剂Cyclo-RGDfK对纤连蛋白诱导乳腺上皮细胞间质转化的影响

目的本研究探讨整合素αvβ3抑制剂(Cyclo-RGDfK)对纤连蛋白(fibronectin,FN)诱导人乳腺上皮细胞MCF-10A上皮间质转化的影响。方法体外传代培养MCF-10A细胞,采用FN诱导建立上皮间质转化(EMT)模型。Western blot实验检测αv integrin、β3 integrin、上皮标志物E-cadherin和间充质标志物N-cadherin、Vimentin的蛋白表达;划痕修复实验和Transwell小室实验检测细胞的迁移和侵袭能力。结果 Western blot结果显示FN作用48 h后可明显增强αv integrin、β3 integrin、N-cadherin、Vimentin的蛋白表达,下调E-cadherin蛋白表达,在给予Cyclo-RGDfK作用后,FN诱导的N-cadherin和Vimentin表达上调和E-cadherin表达下调被明显逆转;划痕修复实验和Transwell小室实验结果表明,FN能显著增强MCF-10A细胞迁移和侵袭能力,Cyclo-RGDfK能显著抑制FN的诱导作用。结论 Cyclo-RGDfK能抑制FN诱导乳腺上皮细胞MCF-10A的EMT过程,降低细胞的侵袭和迁移能力,其可能是治疗乳腺癌转移的潜在药物。

Effects of integrinαvβ3 inhibitor Cyclo-RGDfK on epithelial-mesenchymal transition of breast epithelial cells induced by fibronectin

Aim To investigate the effect of integrin αvβ3 inhibitor(Cyclo-RGDfK)on fibronectin(FN)-induced epithelial-mesenchymal transition in human mammary epithelial cells MCF-10A.Methods MCF-10A cells were cultured in vitro,and the model of epithelial-mesenchymal transition(EMT)induced by FN was established.The expressions ofαv integrin,β3 integrin,epithelial marker E-cadherin and mesenchymal markers N-cadherin and Vimentin were measured by Western blot.The migration ability and invasion ability of cells were detected by Scratch repair experiment and Transwell chamber experiment.Results Western blot results showed that the expressions ofαv integrin,β3 integrin,N-cadherin,and Vimentin significantly were up-regulated and the expression of E-cadherin protein down-regulated after FN treatment for 48 h.After administration of Cyclo-RGDfK,the increased expression of N-cadherin and Vimentin and decreased expression of E-cadherin induced by FN were significantly reversed.Similarly,the results of Scratch repair experiments and Transwell experiments showed that FN significantly enhanced the ability of MCF-10A cells to migrate and invade,and Cyclo-RGDfK significantly inhibited the induction of FN.Conclusions Cyclo-RGDfK can inhibit the process of EMT induced by FN and reduce the invasion and migration of MCF-10A,which may be a potential drug for the treatment of breast cancer metastasis.