【摘要】 目的:探讨转染PSilencer4.1-NET-1 shRNA进入肺癌细胞株A549后对细胞中NET-1基因的表达及细胞增殖的影响。方法:转染PSilencer4.1-NET-1 shRNA进入肺癌细胞株A549,同时设立空转染对照组与空白组,Western blot检测细胞NET-1蛋白,逆转录聚合酶链反应(RT-PCR)检测癌细胞中NET-1mRNA的含量,WST-8法检测细胞的增殖,流式细胞仪检测细胞周期的相关变化。结果:PSilencer4.1-NET-1 shRNA转染组中NET-1 mRNA的含量及细胞NET-1蛋白与对照组比较明显降低,实验组细胞的增殖明显慢于对照组与空白组,癌细胞阻滞在G1期占63.24%,S期细胞比例明显下降,与对照组比较均有统计学意义(P<0.05)。结论:肺癌细胞中NET-1持续低表达可导致癌细胞增殖抑制、生长减慢,在肺癌患者的预后判断中有一定的参考价值。更多还原

【Abstract】 Objective: To explore the effect of PSilencer4.1-NET-1 shRNA on the expression of net-1 gene and proliferation of lung cancer A549 cells.Methods:PSilencer4.1-NET-1 shRNA was transfected into lung cancer A549 cells by inhibiting expression of NET-1 mRNA,and those treated with empty plasmid(control group) and without any treatment(blank group) were compared.The expression of NET-1 mRNA and protein among the stable transfected cells and the untransfected ones was assessed by Western blotting and real-time PCR respectively.The cell growth was examined by Cell Counting Kit-8(MTT) test and cells counted.Finally,cell cycle distribution and cell proliferation were measured by flow cytometry.Results: The expression levels of NET-1 gene mRNA and protein in PSilencer4.1-NET-1 shRNA group were significantly lower compared with the untreated group and controlgroup(P <0.05).In the experimental group,cell proliferation was slower than the control group and the untreated group;63.24% cells were blocked in G 1 phase,nd S phase cells decreased significantly compared with the untreated group and control group(P < 0.05).Conclusion: Sustained low NET-1 expression in lung cancer cells inhibits the proliferation,and shows decreased growth of lung cancer cells.It has some values to judge the prognosis of lung cancer patients.更多还原