Sam68基因过表达可促进乳腺癌MCF-7细胞发生上皮-间质转化

Sam 68 gene-overexpressing accelerates epithelial-mesenchymal transition of breast cancer MCF-7 cells

目的 :探讨Sam68(Src-associated in mitosis 68 k D)对乳腺癌MCF-7细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)、增殖、迁移和侵袭的影响。方法 :将重组质粒pc DNA-3-Sam68转染至乳腺癌MCF-7细胞后,应用实时荧光定量PCR法和蛋白质印迹法分别检测Sam68以及EMT标志物E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和波形蛋白(vimentin)m RNA及蛋白的表达,CCK-8法和Transwell小室法分别检测细胞的增殖、迁移和侵袭情况。结果:重组质粒pc DNA-3-Sam68转染MCF-7细胞48 h后,Sam68m RNA和蛋白明显过表达(P值均<0.05)。EMT标志物E-cadherin、N-cadherin和vimentin m RNA的表达水平无明显变化(P值均>0.05),E-cadherin蛋白的表达水平明显下调(P<0.05),而N-cadherin和vimentin蛋白的表达水平则明显上调(P值均<0.05)。并且,细胞的增殖、迁移和侵袭能力明显增强(P值均<0.05)。结论 :过表达Sam68基因可增强乳腺癌MCF-7细胞的增殖、迁移和侵袭能力,这一作用可能与调控EMT的发生有关。

Objective： To investigate the effects of Src-associated in mitosis 68 kD （Sam68） on the epithelial-mesenchymal transition （EMT）, proliferation, migration and invasion of breast cancer MCF-7 cells. Methods： After the recombinant plasmid of pcDNA-3-Sam68 was transfected into breast cancer MCF-7 cells, the expression levels of Sam68 and EMT markers E-cadherin, N-cadherin and vimentin mRNAs and proteins were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The proliferation, migration and invasion of MCF-7 cells after Sam68 gene-overexpression were detected by CCK-8 method and Transwell chamber assay, respectively. Results： Sam68 mRNA and protein were over-expressed in MCF-7 cells after transfection with pcDNA-3-Sam68 plasmid for 48 h （both P 〈 0.05）. The expression levels of EMT markers E-cadherin, N-cadherin and vimentin mRNAs in MCF-7 cells after transfection with pcDNA- 3-Sam68 plasmid had no changes （all P 〉 0.05）, whereas the expression level of E-cadherin protein was down-regulated （P 〈 0.05）, the expression levels of N-cadherin and vimentin proteins were opposite （both P 〈 0.05）. The proliferation, migration and invasion of MCF-7 cells after transfection with pcDNA-3-Sam68 plasmid were promoted （all P 〈 0.05）. Conclusion： Sam68 and invasion. It is gene-overexpression can accelerate the abilities of proliferation, migration maybe related to regulating the EMT.