目的：研究c-Met对三阴性乳腺癌细胞株MDA-MB-231活力及对阿霉素耐药性的影响。方法：构建阿霉素耐药的MDA-MB-231／ADR细胞系，实时荧光定量PCR及Western blotting技术检测不同细胞系中c-Met mRNA及蛋白的表达。脂质体转染c-Met-siRNA及表达质粒或AKT-siRNA，Western blotting检测转染效率；四甲基偶氮唑法（ MTT法）检测细胞的活力及对阿霉素的敏感性。结果：对阿霉素耐药的MDA-MB-231／ADR细胞中c-Met的mRNA及蛋白表达均显著高于对照的MDA-MB-231细胞，转染高表达c-Met的质粒可增加MDA-MB-231细胞的活力并降低其对阿霉素的敏感性，而利用siRNA抑制耐药细胞株中c-Met的表达后，可以逆转MDA-MB-231／ADR细胞对阿霉素的耐药。此外，c-Met可以磷酸化激活细胞中的AKT，并通过该信号分子增加MDA-MB-231细胞活力并诱导耐药。结论：c-Met可作为一个重要的靶点应用于三阴性乳腺癌的治疗。%[ ABSTRACT] AIM:To investigate the effects of c-Met on the proliferation and the sensitivity to chemotherapeutic drugs of triple negative breast cancer cells.METHODS: Doxorubicin-resistant cells ( MDA-MB-231/ADR) were estab-lished.The expression of c-Met at mRNA and protein levels in the MDA-MB-231/ADR cells and parental MDA-MB-231 cells was detected by real-time PCR and Western blotting.c-Met siRNA and plasmid or AKT siRNA were transfected into the cancer cells.The cell proliferation and the sensitivity to doxorubicin were determined by MTT assay.RESULTS:The expression of c-Met at mRNA and protein levels in MDA-MB-231/ADR cells was significantly higher than that in parental MDA-MB-231 cells.Transfection with pBABE-puro TPR-MET plasmid into the MDA-MB-231 cells induced cell prolifera-tion and resistance to doxorubicin.Meanwhile, inhibition of c-Met in the MDA-MB-231/ADR cells by siRNA reversed the doxorubicin-resistance.In addition, over-expression of c-Met led to higher phosphorylation level of AKT, which was in-volved in the effects of c-Met on the MDA-MB-231 cell proliferation and doxorubicin-resistance.CONCLUSION: c-Met may have the potential as a therapeutic target in the treatment of triple negative breast cancer.
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