KLF5介导p27kip1表达对胶质瘤U87细胞增殖和侵袭的影响

Effects of KLF5-mediated p27kip1 expression on proliferation and invasion of glioma U87 cells

目的观察Kruppel样因子5(KLF5)介导p27kip1对胶质瘤U87细胞增殖和侵袭的影响。方法常规培养胶质瘤细胞U87,将其分为si-NC组和si-KLF5组,si-NC组转染对照siRNA,si-KLF5组转染KLF5 siRNA。采用MTS法检测两组细胞增殖能力,流式细胞术检测细胞周期,Boyden实验检测细胞侵袭能力,Western blotting法检测细胞中细胞周期相关蛋白Cyclin D1、CDK4、CDK6和上皮间质转化蛋白Snail1表达。Western blotting检测显示p27kip1在si-KLF5组的表达水平高于si-NC组,故设置si-KLF5+si-p27kip1组和si-NC组、si-KLF5组,si-KLF5+si-p27kip1组给予p27kip1 siRNA与KLF5 siRNA共同转染U87细胞,si-KLF5组转染KLF5 siRNA,si-NC组转染对照siRNA。比较三组细胞的增殖能力和侵袭能力。结果与si-NC组相比,si-KLF5组U87细胞增殖和侵袭能力均降低(P均<0.05),G1期细胞比例增加(P<0.05),细胞中Cyclin D1、CDK4、CDK6和Snail1蛋白表达降低(P均<0.05)。si-KLF5+si-p27kip1组细胞增殖和侵袭能力均较si-NC组降低(P均<0.05),较si-KLF5组增加(P均<0.05)。结论干扰KLF5表达后U87细胞的增殖和侵袭能力降低,其机制与上调p27kip1表达有关。

Objective To observe the effects of Kruppel-like factor 5(KLF5)-mediated p27kip1 on the proliferation and invasion of glioma U87 cells.Methods U87 glioma cells were cultured routinely and divided into the si-NC group and si-KLF5 group.The U87 glioma cells in the si-NC group were transfected with control siRNA,and the si-KLF5 group with KLF5 siRNA.MTS experiment was used to detect the proliferation of the two groups,flow cytometry was used to detect cell cycle,Boyden experiment was used to detect cell invasion ability,and Western blotting was used to detect the expression of cell cycle-related proteins Cyclin D1,CDK4,CDK6 and epithelial-mesenchymal transition protein Snail1.Western blotting showed that the expression level of p27kip1 in the si-KLF5 group was higher than that in the si-NC group,so the si-KLF5+si-p27kip1 group,the si-NC group,and the si-KLF5 group were set,and the cells in the si-KLF5+si-p27kip1 group U87 cells were co-transfected with p27kip1 siRNA and KLF5 siRNA,the si-KLF5 group with KLF5 siRNA,and the si-NC group with control siRNA,respectively.We compared the proliferation and invasion abilities of the three groups.Results Compared with the si-NC group,the proliferation and invasion abilities of U87 cells in the si-KLF5 group were reduced(both P<0.05),the proportion of cells in the G1 phase increased(P<0.05),and the expression levels of Cyclin D1,CDK4,CDK6 and Snail1 proteins decreased(all P<0.05).The cell proliferation and invasion abilities of the si-KLF5+si-p27kip1 group were lower than those of the si-NC group(all P<0.05),and increased as compared with that of the si-KLF5 group(P<0.05).Conclusion The proliferation and invasion abilities of U87 cells decrease after interference with KLF5 expression,and the mechanism is related to the up-regulation of p27kip1 expression.