沉默LINC00673表达对胶质瘤细胞增殖和侵袭能力的影响及其机制

Effect and mechanism of silencing LINC00673 expression on proliferation and invasion of glioma cells

目的观察沉默LINC00673表达对胶质瘤细胞增殖和侵袭能力的影响,并探讨其可能的分子调控机制。方法选择人神经胶质瘤细胞U251(以下称U251细胞),传代培养。取传5~15代、对数生长期、生长状态良好的U251细胞,随机分为空白对照组、阴性对照组和沉默组,阴性对照组和沉默组按照Lipofectamine2000转染试剂说明分别转染阴性对照siRNA和LINC00673-siRNA。转染48 h,收集细胞,采用CCK-8法检测再培养24、48、72、96 h时各组细胞增殖能力,采用Transwell侵袭实验检测各组细胞侵袭能力,采用Western blotting法检测各组wnt/β-catenin信号通路相关蛋白(wnt、p-wnt、β-catenin、p-β-catenin)表达。结果与空白对照组和阴性对照组比较,沉默组再培养48、72、96 h细胞增殖能力均明显降低,其侵袭细胞数明显减少(P均<0.05),而空白对照组与阴性对照组比较P均>0.05。与空白对照组和阴性对照组比较,沉默组wnt、p-wnt、β-catenin、p-β-catenin蛋白相对表达量均明显降低(P均<0.05),而空白对照组与阴性对照组比较P均>0.05。结论沉默LINC00673表达可抑制胶质瘤细胞增殖和侵袭能力,其机制可能与抑制wnt/β-catenin信号转导通路有关。

Objective To explore the effects of LINC00673 on the proliferation and invasion of glioma cells and the potential mechanism. Methods Glioma cells U251 subcultured for 5-15 times with well condition were selected in this study. U251 cells were randomly divided into three groups： the bank control group,the negative control group,and the knockdown group. Cells in the negative control group and the knockdown group were transfected with the negative controls and LINC00673-siRNA,respectively according to the instrument of the manufacturer of Lipofectamine 2000. At 48 h after transfection,the cells were collected. The proliferation ability was detected by CCK-8 at 24,48,72,and 96 h. The invasion ability was evaluated by Transwell invasion assay. Western blotting was used to detect the expression of wnt/β-catenin pathway related proteins（ wnt,p-wnt,β-catenin,and p-β-catenin）. Results Compared with the bank control group and the negative control group,the proliferation abilities and the number of transmembrane cells in the knockdown group significantly decreased at 48,72,and 96 h（ all P〈0. 05）; the expression of wnt,p-wnt,β-catenin,and p-β-catenin also significantly decreased in the knockdown group（ all P〈0. 05）. However,there was no significant deference in the above parameters between blank control group and negative control group（ all P〉0. 05）. Conclusion Silencing the LINC01133 expression inhibits the proliferation and invasion of glioma cells through inhibiting the wnt/β-catenin pathway.