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影响的研究

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.

Effect of the interaction between Anxa2 and P-gp on migration and invasion of multidrug resistant human breast cancer cells

OBJECTIVE: To investigate the effect of Anxa2 Interacting with P-gp on the migration and invasion of the multidrug resistant(MDR) human breast cancer cells MCF-7/ADR.METHODS: One pair of small interfering RNA(siRNA) targeting P-gp was transfected into MCF-7/ADR cells and monoclonal cell strains were screened.The expression of P-gp was detected by Western blotting.The activity of proliferation was examined by MTT.The migration capacity and the invasion ability were detected by Transwell chambers.The interaction between Anxa2 and P-gp was examined by immunoprecipitation and Immunofluorescence confocal microscopy analysis.RESULTS:Abxa2 and P-gp were expressed in the breast cancer cells.The migration and ivnation of MCF-7/ADR inereased significantly compared with MCF-7.P-gp expression was significantly knocked down and there was a notably decrease in migration and invasion capability of MDR breast cancer cells(P<0.05).There was a close interaction between Anxa2 and P-gp.CONCLUSIONS: Knockdown of P-gp can inhibite migration and invasion capability of MCF-7/ADR cells notably.Moreover,it is discovered that Anxa2 interacted with P-gp by immunoprecipitation and immunofluorescence confocal microscopy analysis indicating that Anxa2 interacting with P-gp may plays an important role in the enhanced invasiveness of the multidrug resistant human breast cancer cells.