上调双特异性磷酸酶2表达对胶质瘤细胞增殖凋亡及STAT3信号通路的影响

Effect of up-regulating DUSP2 on proliferation and apoptosis and STAT3 signaling pathway in the glioma cells

目的:探讨双特异性磷酸酶2(DUSP2)基因对胶质瘤细胞增殖和凋亡的影响及其可能的机制。方法:构建DUSP2过表达慢病毒载体,筛选出DUSP2稳定过表达的胶质瘤细胞株,空载慢病毒转染并筛选后的胶质瘤细胞作为对照。采用CCK8细胞毒性实验和克隆形成实验检测上调DUSP2表达后胶质瘤细胞增殖能力的变化;蛋白质印迹法检测DUSP2、STAT3、p-STAT3(Tyr705/Ser727)、bcl-2、p53等蛋白表达水平。结果:慢病毒载体感染细胞并筛选后,荧光显微镜下观察显示荧光表达效率在90%以上。蛋白质印迹结果显示DUSP2稳定过表达的胶质瘤细胞(实验组)中DUSP2表达较对照组明显上调。CCK8细胞毒性实验结果表明,实验组细胞活力较对照组明显下降;克隆形成实验结果显示,实验组细胞克隆形成数明显少于对照组。蛋白质印迹结果表明,实验组细胞中pSTAT3(Tyr705/Ser727)及bcl-2表达显著下调,p53表达上调。结论:上调DUSP2的表达可显著抑制胶质瘤细胞的增殖,并促进其凋亡,其机制可能与DUSP2抑制了STAT3(Tyr705/Ser727)信号通路的磷酸化水平有关。

Objective： To investigate the effect of dual-specificity phosphatase-2（ DUSP2） on the proliferation and apoptosis in the glioma cells and its mechanisms. Methods： The puromycin-screened DUSP2-overexpression glioma cell line was constructed by the DUSP2-overexpression lentivirus vector,then the ability of proliferation was tested by CCK8 and the cell clone formation assays; the protein expression of DUSP2,STAT3,p-STAT3（ Tyr705/Ser727）,bcl-2 and p53 were tested by the western blotting analysis. Results： After the glioma cells was infected by the lentivirus vector and screened by the puromycin,fluorescence expression efficiency was over 90% under fluorescence microscope. The western blotting analysis revealed that the DUSP2＇s expression of overexpression group（ experimental group） was markedly increased compared with control group. The CCK8 experiment showed that the cell viability of experimental group was significantly decreased than that of the control group. The cell clone formation test showed that the cell clones of the experimental group were obviously fewer than that of the control group. The p-STAT3（ Tyr705/Ser727）,bcl-2 were down regulated and p53 was up regulated in the experimental group. Conclusion： The over-expression of DUSP2 in glioma cells can efficiently inhibit glioma＇s proliferation and promote its apoptosis through the STAT3（ Tyr705/Ser727） signaling pathway.