NET-1 shRNA抑制肺癌细胞A549中NET-1表达及对癌细胞增殖的影响

NET-1 shRNA inhibits expression of net-1 gene and affects proliferation in lung cancer A549 cells

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。

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 canc- er A549 ceils 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 （MTr） 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 G1 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 prolif- eration, and shows decreased growth of lung cancer cells. It has some values to judge the prognosis of lung cancer pa- tients.