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目的研究Rack1分子在TGF-β诱导A549细胞发生上皮-间充质转化(epithelial to mesenchymal transition,EMT)过程中的功能。方法制备携带不同Rack1干涉序列的慢病毒颗粒,并感染A549细胞建立稳定细胞系。选取敲低效率较高的细胞系,利用MTS、细胞划痕、免疫印迹等技术检测Rack1敲低后细胞的增殖、TGF-β诱导的迁移及相关分子表达水平的改变。结果获得了Rack1稳定敲低的细胞系,Rack1敲低后细胞增殖减慢,TGF-β诱导的迁移加快,钙黏蛋白E(E-cadherin)下调和胞外信号调节激酶(extracellular regulated protein kinases,ERK)磷酸化水平升高。结论 Rack1分子敲低促进EMT的发生。
Objective To investigate the role of Rack1 in TGF-β-induced epithelial to mesenchymal transition (EMT) in A549 cells. Methods Lentiviral particles carrying different Rack1 interference sequences were prepared and infected with A549 cells to establish a stable cell line. The knockdown cell lines with high knockdown efficiency were selected and the cell proliferation, the migration induced by Rack-knocked-down and the expression of related molecules were detected by MTS, cell scratch and Western blotting. Results Rack1 stable knockdown cell line was obtained. After Rack1 knockdown, cell proliferation was slowed, TGF-β-induced migration was accelerated, E-cadherin was down-regulated and extracellular regulated protein kinases ERK) phosphorylation level increased. Conclusion Rack1 knockdown can promote the occurrence of EMT.