1MTA2基因沉默对人乳腺癌MCF-7/ADR多药耐药的逆转作用

摘要

目的：乳腺癌是女性中最常见的恶性肿瘤，本研究探讨 MTA2对乳腺癌耐药的作用及机制。方法 CCK-8检测 MCF-7/ADR 和 MCF-7细胞株对阿霉素的耐药情况。体外化学合成 MTA2序列特异性的 shRNA，经慢病毒转染人乳腺癌细胞系，Western blot 检测慢病毒介导的 MTA2敲减效率以及 PI3K/AKT/NF-κB 信号通路蛋白。Annexin V-FITC 和 DAPI 染色检测细胞凋亡情况。结果 MCF-7/ADR 细胞中MTA2表达量显著高于 MCF-7细胞株，慢病毒介导的基因沉默显著的敲减了 MCF-7/ADR 细胞中的 MTA2。敲除 MTA2逆转 MCF-7/ADR 细胞的耐药作用并促进细胞凋亡。敲除 MTA2通过抑制 PI3K/AKT 通路抑制下游 NF-κB 通路活化，并下调 p-gp 蛋白表达而逆转耐药性。结论敲除 MTA2可以逆转 MCF-7/ADR 细胞的耐药作用，表明 MTA2可能参与调节乳腺癌多药耐药作用。%Objective Breast cancer is the most common malignant tumor in women.To investigate the effect and mechanism of MTA2 on breast cancer drug resistance. Methods CCK-8 was used to detect the drug resistance of MCF-7/ADR and MCF-7 cells to adriamycin.MTA2 sequence specific shRNA was synthesized in vitro and transfected into human breast cancer cell line by lentivirus.Knockdown efficiency of lentivirus mediated MTA2 and the signaling pathway protein,such as PI3K,AKT and NF-κB,were detected by Western blot.Apoptosis was detected by Annexin V-FITC and DAPI staining. Results MTA2 expression in MCF-7/ADR cells was significantly higher than that in MCF-7 cell line.Lentivirus-mediated gene silencing significant knockdown MTA2 in MCF-7/ADR cells.Knockdown of MTA2 inhibited the activation of the downstream NF- kappa B pathway by inhibiting the PI3K or AKT pathway,and reversed the resistance to P-gp protein expression. Conclusion Knockout MTA2 can reverse the MCF-7/ADR cells drug resistant effect and these suggest MTA2 may be involved in regulating the role of breast cancer multidrug resistance.