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目的:探讨Anxa2和P-gp蛋白相互作用对多药耐药乳腺癌细胞迁移与侵袭的影响。方法:采用小干扰RNA技术下调人多药耐药乳腺癌细胞MCF-7/ADR中P-gp的表达,并筛选稳定的单克隆细胞株;利用噻唑蓝(MTT)比色法和Transwell实验研究P-gp的表达对MCF-7及其耐药细胞MCF-7/ADR的增殖、迁移和侵袭能力的影响;利用免疫沉淀和免疫荧光分析多药耐药细胞中Anxa2和P-gp的相互关系。结果:蛋白印迹法检测发现,Anxa2和P-gp在耐药乳腺癌细胞中均高表达;MCF-7/ADR与其亲本细胞MCF-7相比,迁移和侵袭能力明显增强;MCF-7/ADR细胞中P-gp的表达被抑制后,其迁移和侵袭能力显著下降,并且差异有统计学意义,P<0.05。免疫沉淀证实,Anxa2和P-gp之间存在相互作用;免疫荧光发现,Anxa2和P-gp在细胞膜上存在很好的共定位。结论:降低P-gp的表达可以明显抑制耐药乳腺癌细胞MCF-7/ADR的迁移和侵袭能力,Anxa2和P-gp之间存在较好的共定位和相互作用。提示Anxa2和P-gp的相互作用有可能对增强多药耐药乳腺癌细胞的侵袭转移能力起到一个重要的作用。
Objective: To investigate the effect of Anxa2 and P-gp protein interactions on the migration and invasion of multidrug-resistant breast cancer cells. Methods: The expression of P-gp in multidrug-resistant human breast cancer cell line MCF-7 / ADR was down-regulated by small interfering RNA technology and the stable monoclonal cell line was screened. MTT assay and Transwell assay P-gp on the proliferation, migration and invasion ability of MCF-7 and its multidrug-resistant cell MCF-7 / ADR; the relationship between Anxa2 and P-gp in multidrug-resistant cells was analyzed by immunoprecipitation and immunofluorescence . Results: Western blotting showed that both Anxa2 and P-gp were highly expressed in the resistant breast cancer cells. Compared with MCF-7, the migration and invasion ability of MCF-7 / ADR was significantly increased. MCF-7 / ADR After the expression of P-gp was inhibited, its migration and invasion ability significantly decreased, and the difference was statistically significant (P <0.05). Immunoprecipitation confirmed the interaction between Anxa2 and P-gp. Immunofluorescence showed that Anxa2 and P-gp had good co-localization on the cell membrane. CONCLUSION: Decreasing the expression of P-gp can significantly inhibit the migration and invasion of MCF-7 / ADR in breast cancer cells. There is a good co-localization and interaction between Anxa2 and P-gp. It is suggested that the interaction between Anxa2 and P-gp may play an important role in enhancing the invasion and metastasis of multidrug-resistant breast cancer cells.