摘要
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 The present study is to investigate IL-24 gene （Ad5F35-hIL-24） effect on the topoisommerase Ⅱα（topo Ⅱα） and Caspase-3 expression in glioma cell line U251. Methods After transfected the U251 glioma cells with the Ad5F35-hIL-24, the methyl thiazolyl tetrazolium （MTT） was used to analyse the inhibition rate of Ad5F35-hIL-24 on the cells. Hoechst 33258 fluorescent staining and flow cytometric assay were used to detect apoptosis. The immunohistoehemistry assay was used to detect topo Ⅱα expression, and Western blotting was applied to detect the protein expression of topo Ⅱα and caspase-3. Transwell experiment was used to test the invasiveness of the cells. Results It was found that the Ad5F35-hIL-24 could inhibit U251 cell proliferation and induce apoptosis in a dose dependent manner compared with the control groups. It showed that Ad5F35-hIL-24 could inhibit topo Ⅱα expression reveled by immunohistochemistry and Westeren blotting, while it increased caspase-3 protein expression. The Transwell experiment showed that the Ad5F35-hIL-24 could reduce the invasiveness of the U251 glioma cells. Conclusion The exogenous IL-24 gene can inhibit the cell proliferation and induce apoptosis of U251 glioma cells. The topo Ⅱα and Caspase-3 are the important molecular targets of the IL-24 gene. These results may give support for the IL-24 gene usage in clinical treatment for glioma patients.
出处
《解剖学报》
CAS
CSCD
北大核心
2010年第1期48-52,共5页
Acta Anatomica Sinica
