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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 effect of Sam68 (Src-associated in mitosis 68 kD) on epithelial-mesenchymal transition (EMT), proliferation, migration and invasion of breast cancer MCF-7 cells. Methods: The recombinant plasmid pcDNA-3-Sam68 was transfected into breast cancer MCF-7 cells. The expression of Sam68 and E-cadherin, the EMT marker, were detected by real-time fluorescence quantitative PCR and Western blot respectively. The expression of N-cadherin and vimentin m RNA and protein were detected. The proliferation, migration and invasion of cells were detected by CCK-8 assay and Transwell chamber assay. Results: The mRNA and protein of Sam68m were overexpressed in MCF-7 cells transfected with recombinant plasmid pcDNA-3-Sam68 for 48 h (P <0.05). The expression of E-cadherin, N-cadherin and vimentin m RNA in EMT markers had no significant difference (P> 0.05), and the expression of E-cadherin protein was significantly decreased (P <0.05) The level of expression was significantly increased (P values <0.05). In addition, the ability of cell proliferation, migration and invasion were significantly enhanced (P <0.05). Conclusion: Overexpression of Sam68 gene can enhance the proliferation, migration and invasion of breast cancer MCF-7 cells, which may be related to the regulation of EMT.