TGF-β1 shRNA对人涎腺黏液表皮样癌Ms细胞生长及粘附能力的影响  

Effects of TGF-β1 shRNA in proliferation and adhesion of human salivary gland mucoepidermoid carcinoma cell Ms in vitro

作　　者：王静[1] 陈健[2] 吴军正 马铭[1] 赵媛[1] 

机构地区：[1]兰州大学口腔医学院,甘肃兰州730000 [2]兰州大学第一医院,甘肃兰州730000 [3]第四军医大学口腔医院,陕西西安710032

出　　处：《癌变．畸变．突变》2010年第1期1-5,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金(30371551);兰州大学医学基金(LZUYX200707)

摘　　要：目的:研究TGF_β1shRNA对人涎腺黏液表皮样癌脊髓转移株Ms(metastatic cells of mucoepidermoid carcinoma in spinal cord,MS)细胞生长及粘附的影响。方法:体外培养Ms细胞、稳定转染空载体的Ms细胞、稳定转染TGF_β1shRNA的Ms细胞,利用细胞计数法,克隆形成实验检测细胞增殖能力的变化;同质粘附实验与异质粘附实验测定细胞的粘附率;考马斯亮蓝染色法检测细胞骨架的改变及免疫荧光法检测Ezrin蛋白的表达。结果:稳定转染TGF_β1shRNA的Ms细胞、稳定转染空载体的Ms细胞和Ms细胞的群体倍增时间分别为30.8,28.8和29.0h;平板克隆形成率分别为14.2%,24.5%和25.1%。Ms细胞转染了TGF_β1shRNA后,30min同质粘附率升高了18.6%;细胞在FN基质表面上1h的粘附抑制率为35.3%。与未转染组、转染空载体组相比,转染TGF_β1shRNA组细胞生长缓慢(P<0.05),同质粘附能力增强(P<0.05),异质粘附能力降低(P<0.05),细胞骨架明显改变,Ezrin蛋白表达降低。结论:TGF_β1shRNA能影响Ms细胞在体外的生长和粘附能力,其机制可能与细胞骨架和Ezrin蛋白表达的改变有关。OBJECTIVE： To study the effects of TGF-β1 shRNA in proliferation and adhesion of human salivary gland mucoepidermoid carcinoma cell Ms in vitro. METHODS： Ms cells, Ms cells transfected with empty vectors and Ms cells transfected with TGF-β1 shRNA were cultured in vitro. Cell count assay, clone formation assay were performed to measure the proliferation of Ms cells. Adhesive abilities were assessed by cell-to-cell adhesion and cell-to-FN adhesion. Coomassie brilliant blue staining assay was used to detect the changes in cytoskeleton and immunoflurescence was used to evaluate the expression of Ezrin protein on Ms cells. RESULTS： The population doubling times of TGF-β1 shRNA-transfected cells, empty vector-transfected cells and the control cells were 30.8, 28.8 and 29.0, respectively. The rates of clone formation were 14.2%, 24.5% and 25.1%, respectively. TGF-β1 shRNA transfection increased the cell-to-cell adhesion by 18.6% in a 30-min adhesion assay, but inhibited the cell-to-FN adhesion by 35.3% in a 1-h adhesion assay. Compared with control and Ms transfected with empty vectors, Ms cells stably transfected with TGF-β1 shRNA grew slowly. The ability of homogeneous adhesion was increased and that of heterogeneous adhesion was inhibited. TGF-β1 shRNA changed the cytoskeleton and down-regulated the expression of Ezrin protein. CONCLUSION： TGF-β1 shRNA could affect the proliferation and adhesion, which may be associated with the changes of cytoskeleton and Ezrin protein expression.

关 键 词：TGF-Β1 SHRNA 黏液表皮样癌 增殖 粘附 

分 类 号：R782.4[医药卫生—口腔医学]