CAJ | 학술논문

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;Hoechst 33258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.
목적 탐토선병독개도적IL-24기인(Ad5F35-hIL-24)대효질류세포계U251탁복이구매Ⅱα(topoⅡα)급Caspase-3표체적영향.방법 응용선병독재체장IL-24기인전염U251세포후,채용사갑기우담서염(MTT)방법관찰Ad5F35-hIL-24대U251세포적억제작용;Hoechst 33258형광염색,이급류식세포술측정세포조망;응용면역조직화학방법검측topoⅡα표체;면역인적법검측topoⅡα화Caspase-3단백질표체변화;Transwell실험관찰Ad5F35-hIL-24대U251세포침습력적영향.결과 여대조조상비,Ad5F35-hIL-24대효질류세포유명현억제작용,능현저유도세포조망,차정농도의뢰성;면역조직화학법현시,Ad5F35-hIL-24능명현억제topoⅡα표체;면역인적검측표명,topoⅡα표체명현강저,이Caspase-3단백적표체수평증가;Transwell실험표명,Ad5F35-hIL-24능명현강저U251세포적침습능력.결론 외원성IL-24기인능현저억제효질류세포증식,유도세포조망;topoⅡα급Caspase-3시기중요적작용파점.해결과대우IL-24기인용우림상치료효질류유일정삼고의의.
Objective The present study is to investigate IL-24 gene(Ad5F35-hIL-24) effect on the topoisommeraseⅡα(topoⅡα) 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 immunohistochemistry assay was used to detect topoⅡα expression, and Western blotting was applied to detect the protein expression of topoⅡα 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 topoⅡα 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 topoⅡα 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.