重组人IL-24蛋白诱导肝癌细胞生长抑制和凋亡  

Growth suppression and apoptosis of liver cancer cells induced by recombined human IL-24 protein(rhIL-24)

作　　者：王翠红[1] 康晓燕[1] 杨之斌[1] 李瑾[1] 钱海华[1] 殷正丰[1] 

机构地区：[1]第二军医大学东方肝胆外科医院分子肿瘤实验室,上海200438

出　　处：《实用肿瘤杂志》2007年第1期18-22,共5页Journal of Practical Oncology

基　　金：国家自然科学基金(30500477);国家973计划项目(2002CB513100)

摘　　要：目的研究重组人IL-24蛋白(rhIL-24)选择性诱导肝癌细胞生长抑制和凋亡的作用。方法重组由Igk前导肽、IL-24 cDNA(不含自身信号肽)和myc-tag组成的融合基因(Igk7m),构建表达载体pcDNA3.1-Igk7m,稳定转染293细胞,表达并粗提分泌性rhIL-24蛋白。以人肝癌细胞株PLC/PRF/5(p53突变型)、SMMC-7721(p53野生型)和正常人肝细胞株WRL-68为靶细胞,用活细胞计数法和TUNEL法分析rhIL-24处理对培养的靶细胞生长和凋亡的影响。结果经测序鉴定,融合基因各片段的核苷酸序列和总阅读框架完全正确。应用Western blot在筛选的转基因293细胞克隆和其培养上清中均检测到rh IL-24蛋白,分子量分别约为25 kDa和35 kDa。与WRL-68对照组相比,分泌性rhIL-24蛋白粗提液处理可显著抑制PLC/PRF/5和SMMC-7721肝癌细胞的生长增殖(P<0.01)。rhIL-24蛋白处理还可显著增加PLC/PRF/5和SMMC-7721的凋亡细胞百分率(P<0.01),而对正常肝细胞WRL-68无明显影响(P>0.05)。结论rhIL-24蛋白能选择性诱导培养的肝癌细胞株PLC/PRF/5和SMMC-7721生长抑制和凋亡。Objective To study the effect of recombined human IL-24 protein （rhIL-24） of seleotively inducing growth suppression and apoptosis on liver cancer cells. Methods To construct a confluent gene （Igk7m） by recombining IL-24 cDNA （signal peptide sequence excluded） with Igk leading peptide sequence and myc-tag. Following stable transfection with pcDNA3.1-IgkTm constructed by cloning IgkTm into pcDNA3. 1 （＋） via Lipofectin, rhIL-24 protein was expressed by 293 cells and distilled rudely. Liver cancer cell lines PLC/PRF/5 （mutate p53）, SMMC-7721 （wild p53） and normal liver cell line WRL-68, were selected to be target cells. After treated with rhIL-24 protein in vitro, viable cells counting and TUNEL were used to analyze influence of rhIL-24 protein on growth and aproptosis of cultured target cells. Results Sequence detection demonstrated the correct nucleotide sequence and total reading frame of each part of confluent gene Igk7m. rhIL-24 protein was detected both in cultured media and inside of transfected 293 cells by Western blot, the molecule weight of each being 35kDa and 25kDa respectively. Contrasted to control group of WRL-68, the secreted rhIL-24 protein rudely distilled could not only notably inhibit growth and proliferation of SMMC-7721 and PLC/PRF/5 cells （P〈0. 01） ,but also increase apoptosis percent of SMMC-7721 and PLC/PRF/5 cells （P〈0. 01）. But as normal liver cells were concerned, rhIL-24 had no notable influence on it （P〉 0. 05）. Conclusion RhIL-24 protein could selectively induce growth suppression and apoptosis of cultured liver cancer cell lines PLC/PRF/5 and SMMC-7721.

关 键 词：肝肿瘤 肿瘤细胞 培养的 重组蛋白质类 凋亡 

分 类 号：R735.7[医药卫生—肿瘤] R73-36[医药卫生—临床医学]