Rab23在脑胶质瘤细胞中的表达及其对细胞增殖的影响

Expression of Rab23 in glioma cell lines U251 and its effect on cell proliferation

目的探讨Rab23在胶质瘤U251细胞系中的表达及其对细胞增殖的影响。方法培养胶质瘤U251细胞系,将细胞分为3组:空白对照组、Rab23转染组和小分子干扰RNA片段(siRNA)干扰组。Rab23转染组脂质体转染Rab23质粒过表达Rab23,siRNA干扰组转染siRNA干扰Rab23的表达。用免疫印迹法测定Rab23蛋白表达水平,实时荧光定量PCR法测定Rab23基因表达水平,噻唑蓝比色法测定干预后U251细胞的增殖水平。结果空白对照组、Rab23转染组和siRNA干扰组的Rab23蛋白表达水平(灰度值)分别为0.82±0.12,1.05±0.23和0.72±0.13,Rab23转染组与空白对照组比较,差异有统计学意义(P<0.01);siRNA干扰组与Rab23转染组比较,差异有统计学意义(P<0.01);在接种第3天,空白对照组、Rab23转染组和siRNA干扰组的细胞增殖水平(OD值)分别为0.75±0.13,0.64±0.05和1.36±0.18,Rab23转染组与空白对照组比较,差异有统计学意义(P<0.01);siRNA干扰组与Rab23转染组比较,差异有统计学意义(P<0.01)。Rab23基因结果的趋势与蛋白一致。结论Rab23在胶质瘤U251细胞中有表达并且该表达对细胞增殖有抑制作用。

Objective To investigate the expression of Rab23 in glioma U251 cell line and its effect on cell proliferation.Methods The glioma U251 cell line was cultured.The cells were divided into three groups:blank control group,Rab23 transfection group and a small interfering RNA fragment(siRNA)interference group.Rab23 transfected liposome transfected Rab23 plasmid overexpression Rab23,and siRNA interference group transfected siRNA interfered with Rab23 expression.The expression level of Rab23 protein was determined by Weston blotting.The expression level of Rab23 gene was determined by real-time PCR.The inhibitory effect of Rab23 on the proliferation of U251 cells was determined by MTT.Results The protein expression of Rab23 protein(grey value)in the control group,Rab23 transfection group and siRNA interference group were 0.82±0.12,1.05±0.23,0.72±0.13,respectively;comparison between Rab23 transfection group and blank control group,the difference was significantly(P<0.01);comparison between siRNA interference group and Rab23 transfection group,the difference was significantly(P<0.01).On the third day of inoculation,the cell proliferation level(OD value)in blank control group,Rab23 transfection group and siRNA interference group were 0.75±0.13,0.64±0.05,1.36±0.18;comparison between Rab23 transfection group and blank control group,the difference was significantly(P<0.01);comparison between siRNA interference group and Rab23 transfection group,the difference was significantly(P<0.01).The trend of gene results is consistent with that of protein.Conclusion Rab23 is expressed in glioma U251 cells and this expression has an inhibitory effect on cell proliferation.