Ad-ING4-IL-24对骨肉瘤生长抑制效应的实验研究  

Experimental study of adenovirus-mediated ING4 and IL-24 gene co-expression on growth suppression of osteosarcoma cells

作　　者：李正祎[1] 缪竞诚[2] 盛伟华[2] 韩亚丽[2] 庄文卓[2] 井莹莹[2] 杨吉成[2] 

机构地区：[1]吉林医药学院检验系,132013 [2]苏州大学医学部细胞与分子生物学教研室,215123

出　　处：《实用医学杂志》2014年第3期349-353,共5页The Journal of Practical Medicine

基　　金：吉林省教育厅"十二五"科学技术研究课题(编号:2012494)

摘　　要：目的:研究肿瘤生长抑制因子4(ING4)和人白细胞介素24(hIL-24)双基因共表达腺病毒载体对MG-63人骨肉瘤细胞及其裸鼠移植瘤的抑癌增效作用。方法:将Ad-ING4-IL-24腺病毒感染MG-63细胞,用RT-PCR法检测ING4和(或)IL-24基因在肿瘤细胞中的表达。MTT法和流式细胞仪(FCM)检测ING4和(或)IL-24基因的表达对MG-63的生长抑制和促凋亡的影响。用骨肉瘤细胞的裸鼠移植瘤动物模型,检测Ad-ING-IL-24对移植瘤生长的抑制作用,并通过免疫组化法检测Bcl-2、Bax、Caspase-3等基因的表达。结果:ING4和IL-24基因在MG-63细胞中成功表达,并对MG-63增殖有明显抑制作用,并能引起细胞凋亡,其凋亡机制可能与Bax/Bcl-2比值升高有关。动物实验表明Ad-ING4-IL-24能显著抑制移植瘤生长,瘤重抑瘤率达75.7%。免疫组化结果显示Ad-ING4-IL-24能明显上调Bax、P21和Caspase-3等基因的表达,同时下调Bcl-2和CD34的表达。结论:Ad-ING4-IL-24在肿瘤基因治疗中是非常理想的抑癌增效载体。Objective To study the enhanced carcinorma-inhibition efficacy of adenovirus-mediated ING4 and IL-24 gene co-expression on growth suppression of osteosarcoma MG-63 cells. Methods The human osteosarcoma MG-63 cells were infected by Ad-ING4-IL-24. The expression level of ING4 and IL-24 were detected by RT-PCR. The growth inhibition and apoptosis effect were detected by MTF, flow cytometry respectively. In athymic nude mice bearing osteosarcoma MG-63 cells, growth inhibition of Ad-ING4-IL-24 on osteosarcoma were measured, the expression of Bcl-2, Bax, Caspase-3, genes expression were tested by immumohistochemistry. Results ING4 and IL-24 genes successfully transcribed in MG-63 cells and could obviously inhibit proliferation and induce cell apoptosis of MG-63, possibly related with Bax/Bcl-2 ratio increasing. Ad-ING4-IL-24 suppressed the tumor growth obviously with an inhibitory rate of 75.7%, obviously up-regulating Bax, P21 and Caspase-3 genes, down-regulating Bcl-2 and CD34 expressions. Conclusion Ad-ING4-IL-24 is a very ideal vector with enhanced anti-tumor effect in gene therapy for tumor.

关 键 词：骨肉瘤 ING4基因 IL-24基因 腺病毒载体 MG-63人骨肉瘤细胞 肿瘤抑制 

分 类 号：R738.1[医药卫生—肿瘤]