CAJ | 학술논문

目的:研究TOF-β1 shBNA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-β1 shRNA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-β1 shRNA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29.0 h;平板克隆形成率分别为14.2%,24.5%和25.1%.Ms细胞转染了TGF-β1 shRNA后,30 min同质粘附率升高了18.6%;细胞在FN基质表面上1 h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-β1 shRNA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-β1 shRNA能影响Ms细胞在体外的生长和枯附能力,其机制可能与细胞骨架和Ezrin蛋白表达的改变有关.
목적:연구TOF-β1 shBNA대인연선점액표피양암척수전이주Ms(metastatic cells of mucoepidermoid carcinoma in spinal cord,MS)세포생장급점부적영향.방법:체외배양Ms세포、은정전염공재체적Ms세포、은정전염TGF-β1 shRNA적Ms세포,이용세포계수법,극륭형성실험검측세포증식능력적변화;동질점부실험여이질점부실험측정세포적점부솔;고마사량람염색법검측세포골가적개변급면역형광법검측Ezrin단백적표체. 결과:은정전염TGF-β1 shRNA적Ms세포、은정전염공재체적Ms세포화Ms세포적군체배증시간분별위30.8,28.8화29.0 h;평판극륭형성솔분별위14.2%,24.5%화25.1%.Ms세포전염료TGF-β1 shRNA후,30 min동질점부솔승고료18.6%;세포재FN기질표면상1 h적점부억제솔위35.3%.여미전염조、전염공재체조상비,전염TGF-β1 shRNA조세포생장완만(P<0.05),동질점부능력증강(P<0.05),이질점부능력강저(P<0.05),세포골가명현개변,Ezrin단백표체강저.결론:TGF-β1 shRNA능영향Ms세포재체외적생장화고부능력,기궤제가능여세포골가화Ezrin단백표체적개변유관.
OBJECTIVE: To study the effects of TGF-β 1 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. Ccomassie brilliant blue staining assay was used to detect the changes in cyteskeleton and immunoflurescence was used to evaluate the expression of Ezrin protein on MS cells, RESUKTS: 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 central 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 he associated with the changes of cytoskeleton and Ezrin protein expression.