沉默Sam68基因表达抑制乳腺癌细胞的上皮-间质转化  

Sam 68 gene-silencing inhibits epithelial-mesenchymal transition of breast cancer cells

作　　者：邝紫桥[1] 章乐虹[1] 潘小梅[1] 刘毅俊[1] 邹颖[1] 陈欣欣[1] 

机构地区：[1]广州医科大学附属第二医院乳腺外科,广东广州510260

出　　处：《肿瘤》2016年第7期740-748,共9页Tumor

基　　金：广东省科技厅基金项目(编号:2010B031600294);广东省自然科学基金项目(编号:2015A030313462)~~

摘　　要：目的 :探讨沉默Sam68(Src-associated substrated during mitosis of 68KD)基因表达对乳腺癌MDA-MB-231细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)的影响。方法 :采用实时荧光定量PCR和蛋白质印迹法检测具有不同侵袭能力的乳腺癌MDA-MB-231和MCF-7细胞中Sam68 m RNA及蛋白的表达情况;选择Sam68高表达的MDA-MB-231细胞,利用si RNA技术沉默细胞中内源性Sam68蛋白的表达。随后,采用实时荧光定量PCR和蛋白质印迹法检测EMT标志物E-钙黏蛋白(E-cadherin)和波形蛋白(vimentin)m RNA及蛋白的表达;采用CCK-8法、Transwell小室迁移和侵袭实验检测沉默Sam 68基因表达后对乳腺癌MDA-MB-231细胞增殖,迁移和侵袭的影响。结果 :Sam68蛋白在高侵袭性的间质型乳腺癌MDA-MB-231细胞中高表达(P<0.05);采用si RNA沉默MDA-MB-231细胞内源性Sam68蛋白的表达后,E-cadherin的表达水平明显上调(P<0.05),而vimentin的表达水平明显下调(P<0.05);MDA-MB-231细胞的增殖能力以及细胞的迁移和侵袭能力明显降低(P值均<0.05),细胞可能发生间质-上皮转化。结论 :Sam68在高侵袭性乳腺癌细胞中高表达,沉默Sam68基因的表达能抑制EMT的发生,并降低细胞增殖、侵袭和迁移能力。Objective：To investigate the effects of Src-associated substrated during mitosis of 68KD（Sam68） gene-silencing on epithelial —mesenchymal transition（EMT） of breast cancer MDA-MB-231 cells.Methods：The expression levels of Sam68 mRNA and protein in breast cancer MDA-MB-231 and MCF-7 cells with different invasive abilities were detected by real-time fluorescent quantitative PCR and Western blotting,respectively.The breast cancer MDA-MB-231 cells with the higher expression of Sam68 were transfected with siRNA to down-regulate the expression of endogenous Sam68 gene.Then the expression levels of EMT markers E-cadherin（epithelial type） and vimentin（interstitial type）were detected by real-time fluorescent quantitative PCR and Western blotting,respectively.The proliferation,migration and invasion abilities of MDA-MB-231 cells after Sam68 genesilencing were detected by CCK-8 method and Transwell migration and invasion assay,respectively.Results：The expression level of Sam68 protein in breast cancer MDA-MB-231 cells was higher than that in MCF-7 cells（P〈0.05）.After endogenous Sam68 gene-silencing by siRNA transfection,the expression level of E-cadherin protein in MDA-MB-231 cells was significantly up-regulated（P〈0.05）,but the expression level of vimentin was significantly down-regulated（P〈0.05）,while the cell proliferation,migration and invasion abilities were significantly reduced as compared with those of the untransfected group（all P〈0.05）.Conclusion：Sam68 protein is highly expressed in highly invasive breast cancer cells.Sam68gene-silencing can reverse the occurrence of EMT,and reduce the proliferation,invasion and migration abilities of breast cancer cells.

关 键 词：乳腺肿瘤 Sam68基因 上皮-间质转化 RNA 小分子干扰 

分 类 号：R737.9[医药卫生—肿瘤]