抑制STAT3在胶质瘤细胞中的表达导致细胞凋亡

Inhibition of STAT3 Gene Expression in Inducing Apoptosis of Human Glioma Cells Line SHG44

目的探讨Stat3基因在脑胶质瘤细胞中的表达及作用。方法将对应Stat3基因特异序列的发夹结构RNA(small hairpin RNA,shRNA)表达载体转染到脑胶质瘤细胞株SHG44,观察细胞形态的变化;用TdT介导的dUTP缺口末端标记技术(TUNEL)对细胞进行检测;RT-PCR分析Stat3的mRNA的表达量变化。结果转染shRNA表达载体使脑胶质瘤细胞形态发生显著变化;TUNEL实验观察到细胞凋亡数目明显增多,且凋亡细胞有明显荧光;RT-PCR结果显示转染后Stat3 mRNA的表达量明显下降。结论 Stat3蛋白在脑胶质瘤细胞正常生长中起到重要作用,抑制该蛋白在细胞中的表达会导致细胞凋亡。

Stat3 gene-targeting by RNAi provides a useful method to study the function and mechanisms of Glioma,and may pave for gene therapy of the encephalic malignancy of Glioma.Here shRNA expression vector which is formed by a pair of DNA sequences corresponding to stat3 gene was constructed,ligated with pSilenCircle vector containing U6 Promotor of RNA polⅢ.The DNA sequences could transcript small hairpin RNA.Terminal deoxynucleofidyl transferase（TdT）-mediated dUTP nick end-labelling（TUNEL） was used to detect the amounts of apoptosis cells.And RT-PCR assay was used to analyse the expression amounts of stat3 mRNA after transferring shRNA vector into glioma cells.Glioma cell line SHG44 underwent prominent morphological changes after transfection.TUNEL assay indicated that much more cells were apoptosis and had apparent fluorescence than blank constrast group.RT-PCR results shown that there was a reduction of the mRNA level in the treatment group.These findings suggest that stat3 has important function in promoting the natural growth of Glioma,and inhibiting the expression of stat3 will induce apoptosis of Glioma.