IL-24基因重组腺病毒载体转染的树突状细胞促进宫颈癌CaSki细胞凋亡  

Dendritic cells transfected with recombinant adenovirus carrying IL-24 gene induces CTLs to promote apoptosis of cervical cancer cell line CaSki

作　　者：潘巍巍[1] 曹利仙[1] 徐营[1] 沈忠飞[1] 易发平[2] 宋方洲[2] 

机构地区：[1]嘉兴学院医学院生物教研室,浙江嘉兴314001 [2]重庆医科大学生物化学与分子生物学教研室,重庆400016

出　　处：《中国病理生理杂志》2014年第1期41-47,共7页Chinese Journal of Pathophysiology

基　　金：浙江省自然科学基金资助项目(No.LQ13H160020;No.Y2110873;No.Y2110158);嘉兴市科技局科研项目(No.2008AY2039-3;No.2010AY1068;No.2011AY1048-6);"十二五"浙江省高校药理学重点学科资助项目

摘　　要：目的:探讨白细胞介素24(interleukin-24,IL-24)基因重组腺病毒载体转染的树突状细胞(dendritic cells,DCs)是否在体外诱导细胞毒性T淋巴细胞(cytotoxic T-lymphocytes,CTLs)对宫颈癌CaSki细胞的杀伤效应。方法:分离培养小鼠未成熟DCs,将已成功构建的带有IL-24基因的重组腺病毒感染体外培养的小鼠未成熟DCs,提取CaSki细胞裂解物负载DCs,制备DC疫苗,Western blotting检测IL-24蛋白的表达;PE-Annexin V和Western blotting检测细胞凋亡及cleaved caspase-3表达;克隆形成实验检测克隆形成能力;裸鼠荷瘤模型体内研究带有IL-24基因的重组腺病毒载体转染DCs在体外诱导的CTLs对宫颈癌CaSki细胞的杀伤效应。结果:Western blotting检测IL-24蛋白高表达;DCs疫苗诱导产生的CTLs后与CaSki细胞共培养,对CaSki细胞生长具有明显的抑制作用,DCs疫苗诱导产生的CTLs促进CaSki细胞凋亡,增加cleaved caspased-3蛋白表达,克隆形成实验检测该疫苗具有抑制CaSki细胞形成克隆的能力。裸鼠的成瘤实验表明,带有IL-24基因的重组腺病毒载体转染DCs可以抑制体内肿瘤的形成,促进肿瘤细胞凋亡。结论:IL-24基因重组腺病毒感染DCs制备的DCs疫苗可诱导CTLs,从而抑制宫颈癌CaSki细胞增殖,并促进其凋亡。AIM： To investigate the therapeutic potential of interleukin-24 （IL-24） in cancer treatment, we observed the inductive effect of eytotoxic T-lymphocytes （CTLs） against human cervical cancer cell line CaSki using den- dritic cells （DCs） transfected with recombinant adenovirus carrying IL-24 gene （Ad-IL-24-DCs）. METHODS ： Immature mouse DCs were isolated and cultured. The DCs were infected with recombinant adenovirus carrying IL-24 gene. The CaSki cell lysate-loaded autologous DCs vaccine was also prepared. The cell proliferation was analyzed by MTY assay. The cell apoptosis was determined by flow cytometry with PE-Annexin V staining. The cleaved caspase-3 in the Ad-IL-24-DCs was measured by Western blotting. Colony-forming assay was used to detect the colony number of Ad-IL-24-DCs. The tumori- genic capacity of CaSki cells in vivo in the presence of Ad-IL-24-DCs was observed by tumor-burdened model. RESULTS： IL-24 was highly expressed in the Ad-IL-24-DCs. In the presence of Ad-IL-24-DCs, the apoptosis of CaSki ceils was signif- icant increased. Additionally, the protein level of cleaved caspase-3 was increased in CaSki ceils in the presence of Ad-IL- 24-DCs, suggesting that the CaSki cells with Ad-IL-24-DCs induced apoptosis. CTLs induced by Ad-IL-24-DCs inhibited the tumorigenic capacity of CaSKi cells in vivo. CONCLUSION： Genetically modified DCs are successfully prepared by infection with Ad-IL-24 and show a significant effect on triggering specific CTLs against CaSki ceils.

关 键 词：白细胞介素24 腺病毒 树突状细胞 细胞凋亡 

分 类 号：R373.39[医药卫生—病原生物学]