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目的探讨腺病毒介导的IL-24基因(Ad5F35-hIL-24)对胶质瘤细胞系U251拓扑异构酶Ⅱα(topoⅡα)及Caspase-3表达的影响。方法应用腺病毒载体将IL-24基因转染U251细胞后,采用四甲基偶氮唑盐(MTT)方法观察Ad5F35-hIL-24对U251细胞的抑制作用;Hoechst33258荧光染色,以及流式细胞术测定细胞凋亡;应用免疫组织化学方法检测topoⅡα表达;免疫印迹法检测topoⅡα和Caspase-3蛋白质表达变化;Transwell实验观察Ad5F35-hIL-24对U251细胞侵袭力的影响。结果与对照组相比,Ad5F35-hIL-24对胶质瘤细胞有明显抑制作用,能显著诱导细胞凋亡,且呈浓度依赖性;免疫组织化学法显示,Ad5F35-hIL-24能明显抑制topoⅡα表达;免疫印迹检测表明,topoⅡα表达明显降低,而Caspase-3蛋白的表达水平增加;Transwell实验表明,Ad5F35-hIL-24能明显降低U251细胞的侵袭能力。结论外源性IL-24基因能显著抑制胶质瘤细胞增殖,诱导细胞凋亡;topoⅡα及Caspase-3是其重要的作用靶点。该结果对于IL-24基因用于临床治疗胶质瘤有一定参考意义。
Objective To investigate the effect of adenovirus-mediated Ad5F35-hIL-24 on the expression of topoⅡα and Caspase-3 in glioma cell line U251. Methods After transfection of U251 cells with adenovirus vector, the inhibitory effect of Ad5F35-hIL-24 on U251 cells was observed by MTT assay; Hoechst33258 fluorescence staining and flow cytometry The expression of topoⅡα was detected by immunohistochemical method. The expressions of topoⅡα and Caspase-3 protein were detected by Western blotting. The effect of Ad5F35-hIL-24 on invasiveness of U251 cells was observed by Transwell assay. Results Compared with the control group, Ad5F35-hIL-24 significantly inhibited the apoptosis of glioma cells in a concentration-dependent manner. Immunohistochemistry showed that Ad5F35-hIL-24 significantly inhibited topoⅡα The expression of topoⅡα was significantly decreased and the expression of Caspase-3 protein was increased by Western blotting. Transwell assay showed that Ad5F35-hIL-24 significantly reduced the invasion ability of U251 cells. Conclusion Exogenous IL-24 gene can significantly inhibit glioma cell proliferation and induce apoptosis; topoⅡα and Caspase-3 are important targets. The results for the IL-24 gene for clinical treatment of glioma has some reference.