摘要
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 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.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2012年第3期166-171,共6页
Chinese Journal of Cancer Prevention and Treatment
基金
国家自然科学基金(81071731)
教育部长江学者和创新团队发展计划项目(IRT1076)
作者简介
【第一作者简介】张海嫦，女，江西高安人，硕士，主要从事肿瘤发生和发展以及转移分子机制的研究工作。Tel：86—22—23340123 E—mail：zhanghaichang2006@163．com
【通讯作者简介】牛瑞芳，女，山西阳泉人，博士，研究员，教授，硕士生导师，主要从事肿瘤发生和发展以及转移分子机制的研究工作。Tel：86—22--23340123—5549 E—mail：niurf1982@yahoo．com．cn
