YB-1对乳腺癌MCF-7细胞CD44表达和细胞侵袭及成球能力的影响  

Effect of YB-1 on the expression of CD44,invasion ability,and mammosphere formation in breast cancer cells MCF-7

作　　者：吴晓君[1] 许文林[1] 冷加燕[1] 龙璐璐[1] 朱小兰[1] 李皓[1] 姚俊[1] 徐红美[1] 陈巧云[1] 惠璐璐[1] 

机构地区：[1]江苏大学附属人民医院中心实验室,江苏镇江212002

出　　处：《江苏大学学报（医学版）》2011年第4期313-318,F0002,共7页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(39970832);江苏省卫生厅重大课题资助项目(K2005017)

摘　　要：目的:观察Y-盒结合蛋白1(Y-box binding protein-1,YB-1)对乳腺癌MCF-7细胞CD44表达及细胞迁移、成球能力的影响。方法:应用基因重组技术将YB-1基因及其siRNA构建于腺病毒载体pADeasy-1,重组腺病毒及腺病毒空载体转染乳腺癌细胞株MCF-7;通过蛋白质印迹法,RT-PCR和免疫荧光的方法检测YB-1和CD44的表达;MTT法检测转染后细胞生长速度,并绘制生长曲线;并通过Transwell小室法检测细胞侵袭和迁移能力;细胞使用无血清培养基培养,并加入表皮生长因子(EGF)和碱性成纤维细胞生长因子(bFGF)后,观察YB-1对细胞成球能力的影响。结果:蛋白质印迹法和RT-PCR结果显示,转染YB-1重组腺病毒载体的MCF-7细胞中,YB-1和CD44的表达明显高于转染腺病毒空载体组和未转染组;相反,转染YB-1 siRNA重组腺病毒载体的MCF-7细胞中,YB-1和CD44表达明显低于对照腺病毒空载体组和未转染组;免疫荧光也证实CD44蛋白水平与YB-1表达正相关。细胞生长曲线表明,转染YB-1重组腺病毒载体组的细胞生长速度最快,而且其侵袭、迁移和成球能力最强。结论:YB-1可能通过调控CD44的表达影响人乳腺癌细胞MCF-7的侵袭及成球能力。Objective： To explore the effects of YB-1 on regulating CD44 and invasion ability and mammosphere formation of breast cancer cells MCF-7.Methods： YB-1 and its siRNA recombined adenovirus vectors were transfected into MCF-7 cell lines,and the expressions of YB-1 and CD44 were tested after the transfection by RT-PCR and Western blotting.Immunofluorescence was also used to confirm the CD44 protein expression changes.MTT was used to test the tumor cell proliferation and a cell growth curve was drawn afterward.Transwell chamber models were used to test the capabilities of cell invasion and migration.For evaluating the abilities of MCF-7 cell forming mammospheres after the transfection,a sphere growth test was applied by culturing cells in serum-free medium plus EGF（20 ng/ml） and bFGF（10 ng/ml）.Result： According to the results of RT-PCR and Western blotting,the expressions of YB-1 and CD44 were enhanced after MCF-7 cells were transfected with YB-1 recombined adenovirus vectors,compared to the control groups.On the contrary,the expressions of YB-1 and CD44 were reduced by transfecting YB-1 siRNA recombined adenovirus vectors.The results of CD44 in immunofluorescence experiments indicated the same conclusion of RT-PCR and Western blotting.The cell proliferation curve showed that MCF-7 cells transfected with YB-1 recombined adenovirus vectors possessed the best growth abilities,and the cells transfected with YB-1 siRNA recombined adenovirus vectors,as expected,showed the lowest growth speed among all groups.MCF-7 cells introduced with YB-1 recombined adenovirus vectors also had the strongest invasion,migration,and mammosphere formation abilities,while the cells transfected with YB-1 siRNA recombined adenovirus vectors were opposite.Conclusion： High expression of YB-1 might potentially enhance the expression of tumor stem cell gene CD44,in addition,to increase the invasion and mammosphere formation in breast tumor cells.

关 键 词：YB-1 CD44 乳腺癌 MCF-7细胞株 侵袭 肿瘤成球能力 

分 类 号：R73-35[医药卫生—肿瘤]