MiR-133通过RHOA蛋白调节乳腺癌细胞系MCF-7的侵袭转移  

MiR-133 regulating the invasive and migrating ability of breast cancer cell line MCF-7 through RHOA protein

作　　者：墨青青[1] 陈平波[1] 卢运萍[1] 马丁[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,武汉市430030

出　　处：《实用医学杂志》2010年第4期541-543,共3页The Journal of Practical Medicine

基　　金：国家973重点基础科学基金资助项目(编号:2009CB218900);国家自然基金(编号:30600668)

摘　　要：目的:探讨miR-133通过调节RHOA蛋白的表达对乳腺癌低转移细胞系MCF-7侵袭转移能力的影响。方法:(1)将has-miR-133a inhibitor经脂质体包裹转入MCF-7细胞,以Negative control作为阴性对照,未做任何处理的细胞作为空白对照。(2)通过qRT-PCR检测转染has-miR-133a inhibitor后细胞中miR-133的表达情况。(3)Transwell试验检测miR-133对于细胞迁移能力的影响。(4)细胞免疫荧光观察miR-133对于细胞骨架形态的影响。(5)Western blot检测miR-133对于RHOA蛋白表达的影响。结果:(1)转染has-miR-133a inhibitor后细胞中miR-133的表达较对照组明显减低。(2)Transwell试验结果表明抑制miR-133后可以增加细胞的迁移能力。(3)细胞免疫荧光结果提示抑制miR-133后细胞骨架形态发生明显改变,细胞具有较强的侵袭转移特性。(4)Western blot结果提示抑制miR-133后RHOA的表达较两个对照组明显上调。结论:miR-133可以通过调控RHOA蛋白的表达来抑制乳腺癌细胞系MCF-7的侵袭转移。Objective To investigate the effects of miR-133 on the invasive and migrating ability of breast cancer lowly metastatic cell line MCF-7 through regulating the expression of RHOA protein. Methods MCF-7 cells were transfected with has-miR-133a inhibitor by Lipofectamine,MCF-7 cells transfected with Negative control were cultured as negative control,and untreated cells as blank control. The expression of miR-133 in MCF-7 cells transfected with has-miR-133a inhibitor was detected by qRT-PCR. The effects of miR-133 on the migrating ability of MCF-7 cells were detected by Transwell experiment. The effects of miR-133 on skeleton of MCF-7 cells were observed by immunofluorescence. And the effects of miR-133 on the expression of RHOA protein were detected by Western blot. Results The expression of miR-133 in MCF-7 cells transfected with has-miR-133a inhibitor was evidently reduced as compared with controls. Inhibiting the miR-133 can enhance the migrating ability of MCF-7 cells. The skeleton of MCF-7 cells was evidently changed after inhibiting miR-133,and MCF-7 cells had more powerful invasive and migrating ability. And the expression of RHOA protein in MCF-7 cells transfected with has-miR-133a inhibitor was significantly higher than those in the other two controls. Conclusion MiR-133 can suppress the invasion and migration of breast cancer cell line MCF-7 through regulating the expression of RHOA protein.

关 键 词：乳腺肿瘤 MiR-133 RHOA蛋白 侵袭转移 

分 类 号：R737.9[医药卫生—肿瘤]