SUMO蛋白质修饰化CRMP2在胶质瘤细胞中表达变化及对细胞增殖的影响

Expression changes of SUMOylation CRMP2 in glioma cells and its effect on cell proliferation

目的观察SUMO蛋白质修饰化CRMP2在胶质瘤细胞中的表达变化及对细胞增殖的影响。方法培养人胶质瘤细胞系A172、U87、GL15,检测细胞中的Ucb9、CRMP2及SUMO蛋白质修饰化的CRMP2蛋白。选取人胶质瘤细胞系GL15进行后续研究。将人胶质瘤细胞分为空白对照组、pCMV-CRMP2组、pCMV-CRMP2-K374A组,pCMV-CRMP2组、pCMV-CRMP2-K374A组、空白对照组分别转染pCMV-CRMP2、pCMV-CRMP2-K374A(定点诱导突变使细胞被SUMO蛋白质修饰化能力下降)及pCMV质粒载体,采用细胞增殖实验检测三组细胞增殖能力。将胶质瘤细胞分为对照组和Ucb9组,Ucb9组转染Ucb9类似物,对照组转染等量二甲基亚砜,检测两组细胞增殖能力。结果GL15、A172、U87细胞中均表达CRMP2、Ucb9,3种胶质瘤细胞CRMP2、Ucb9表达由高到低依次为U87、GL15、A172细胞,两两相比,P均<0.05。GL15、A172、U87细胞中CRMP2与Ucb9表达均呈正相关(r分别为0.87、0.79、0.81,P均<0.05),SUMO蛋白质修饰化CRMP2与Ucb9表达呈正相关(r分别为0.76、0.80、0.77,P均<0.05)。pCMV-CRMP2组细胞增殖能力高于pCMV-CRMP2-K374A组及空白对照组(P均<0.05)。Ucb9组细胞增殖能力高于对照组(P<0.05)。结论人胶质瘤细胞系A172、U87、GL15均表达Ucb9、CRMP2及CUMO蛋白质修饰化CRMP2,3种胶质瘤细胞中CRMP2表达与Ucb9及SUMO蛋白质修饰化CRMP2表达呈正相关;Ucb9介导CRMP2的SUMO蛋白质修饰化可能对胶质瘤细胞增殖起到促进作用。

Objective To explore the expression changes of SUMOylation CRMP2 in glioma cells and its effect on cell proliferation.Methods We cultured the human glioma cell lines A172,U87,GL15 anddetected Ucb9,CRMP2 and SUMOylation CRMP2 in the cells.The human glioma cell line GL15 was selected for the follow-up experiments.Human glioma cells were divided into the blank control group,pCMV-CRMP2 group,and pCMV-CRMP2-K374A group.The cells in the pCMV-CRMP2 group,pCMV-CRMP2-K374A group,and the blank control group were transfected with pCMV-CRMP2,pCMV-CRMP2-K374A(Use Q5Site-Directed Mutagenesis Kit to induce mutant nucleotides through site-directed mutagenesis,which reduces the ability of glioma cells to be acetylated),and pCMV plasmid vector.The cell proliferation experiment was used to detect the proliferation ability of cells in the three groups.We divided the glioma cells into the control group and Ucb9 group.The cells in the Ucb9 group were transfected with Ucb9 analogues,and the control group with the same amount of DMSO.The cell proliferation experiment was used to detect the cell proliferation ability of the two groups.Results Both CRMP2 and Ucb9 were expressed in GL15,A172,and U87 cells.Among these three glioma cells,the order of CRMP2 and Ucb9 expression from high to low levels was U87,GL15,and A172 cells,and the difference was statistically significant between every two groups(all P<0.05).The expression levels of CRMP2 and Ucb9 were all positively correlated in GL15,A172,and U87 cells(r=0.87,0.79,0.81,respectively;all P<0.05),and the expression of SUMOylation CRMP2 was positively correlated with Ucb9 expression(r=0.76,0.80,0.77,respectively;all P<0.05).The cell proliferation ability of pCMV-CRMP2 group was higher than those of pCMV-CRMP2-K374A group and blank control group(both P<0.05).The cell proliferation ability of the Ucb9 group was higher than that of the control group(P<0.05).Conclusions Human glioma cell lines A172,U87,GL15 all express Ucb9,CRMP2 and SUMOylation CRMP2.The expression of CRMP2 in the three glioma cells is positively correlated with the expression of Ucb9 and SUMOylation CRMP2.Ucb9-mediated SUMOylation of CRMP2 may promote the proliferation of glioma cells.