目的:探讨大鼠C6胶质瘤细胞中GDNF选择性剪接体的表达情况及意义。方法:通过检测C6胶质瘤细胞中GDNF基因△78位点剪接体α-pro-GDNF mRNA和β-pro-GDNF mRNA及其受体蛋白GFRα1、GFRα2、Ret和NCAM表达,结合介导肿瘤迁移相关蛋白RhoA、Cdc42、Rac1的表达情况,分析其相关关系。结果:(1)C6胶质瘤细胞中,可以检测到α-pro-GDNF和β-pro-GDNF的mRNA表达,且α-pro-GDNF mRNA表达水平比β-proGDNF高;与星形胶质细胞相比,α-pro-GDNF mRNA表达水平的增高趋势较β-pro-GDNF明显;(2)C6胶质瘤细胞中RhoA mRNA表达水平与α-pro-GDNF mRNA表达水平成显著正相关(R2=0.699,P〈0.05),Cdc 42 mRNA表达水平与α-pro-GDNF mRNA表达水平呈显著正相关(R2=0.803,P〈0.05),与β-pro-GDNF mRNA表达相关性不显著。结论:C6胶质瘤中α-pro-GDNF具有优先表达,可能与胶质瘤细胞的迁移具有相关性。
Objective: To investigate the expression and significance of GDNF splice variants in rat C6 glioma cells.Methods: Real-time PCR techniques were used to detect the expression level of mRNA of both GDNF splice variants( α-and β-pro-GDNF),GDNF receptors( GFRα1,GFRα2,Ret and NCAM),associated proteins mediated tumor migration( RhoA,Cdc42,Rac1) in normal rat astrocytes and C6 glioma cells,and the relationship between them.Results:( 1) Both α- and β-pro-GDNF mRNA were expressed in C6 glioma cells,and α-pro-GDNF mRNA expression levels were higher than β-pro-GDNF; Compared to astrocytes,the elevated degree of expression levels of α-pro-GDNF mRNA was significantly more than β-pro-GDNF.( 2) The mRNA expression level of RhoA( R2= 0. 699,P〈0. 05) and Cdc42( R2= 0. 803,P〈0. 05) had a significantly positive correlation with α-pro-GDNF mRNA expression level,but no relationship with β-pro-GDNF mRNA expression level. Conclusion: GDNF alternative splicing α-pro-GDNF expressed abuadautly in C6 glioma cells and was possibly correlated with the migration of glioma cells.