亚砷酸联合重组腺病毒Ad-IκBaM对肝细胞癌的治疗作用  

Therapeutic effect of arsenious acid combined with recombinant adenovirus Ad-IκBaM on hepatocellur carcinoma

作　　者：刘丹[1] 刘冰熔[1] 胡丽红[1] 杜雅菊[1] 裴凤华[1] 吕志武[1] 关景明[1] 

机构地区：[1]哈尔滨医科大学附属二院消化内科,黑龙江省哈尔滨市150086

出　　处：《世界华人消化杂志》2006年第33期3169-3174,共6页World Chinese Journal of Digestology

基　　金：黑龙江省科学技术计划项目;No.GC02C148-01~~

摘　　要：目的:观察感染Ad-IκBaM对NF-κB激活的抑制及对亚砷酸诱导肝癌细胞凋亡的增强效应,探讨亚砷酸对肝癌细胞的治疗作用.方法:选择人肝癌细胞系BEL-7402和SMMC-7721,以不同浓度亚砷酸处理.制备重组腺病毒Ad-IκBaM,用来转染经和未经亚砷酸处理的肝癌细胞.MTT和TUNEL方法观察各组细胞生长及凋亡情况;应用EMSA及Westernblot研究肝癌细胞核内NF-κB的激活情况和感染Ad-IκBaM对NF-κB激活的抑制效果.结果:MTT结果表明各浓度亚砷酸对肝癌细胞的作用较正常肝细胞显著(P<0.01);Westernblot和EMSA结果提示亚砷酸可明显抑制肝癌细胞生长,使细胞内NF-κB系统活化;感染重组腺病毒Ad-IκBaM的肝癌细胞后,亚砷酸引起的NF-κB的激活受到明显抑制.亚砷酸联合Ad-IκBaM或Ad-IκBa作用于肝癌细胞SMMC-7721的凋亡指数分别为66.47%和36.67%;亚砷酸联合Ad-IκBaM或Ad-IκBa作用于肝癌细胞Bel-7402的凋亡指数分别为74.5%和32.37%.结论:亚砷酸对肝癌细胞有明显的杀灭作用,同时激活了肝癌细胞内的NF-κB;应用重组腺病毒Ad-IκBaM可以有效抑制NF-κB的激活,并可明显增强亚砷酸对肝癌细胞的作用.AIM： To observe the inhibitory effect of recombinant adenovirus Ad-IkBαM on the activation of nuclear factor kappa B （NF-kB） as well as the enhancing effect of arsenious acid on the apoptosis of hepatocellular carcinoma cells. METHODS： Hepatocellular carcinoma cell lines BEL-7402 and SMMC-7721 were treated with different concentrations of arsenious acid, respectively. The recombinant adenoviruses were prepared to transfect BEL-7402 and SMMC-7721 cells received or not received arsenious acid treatment. MTT assay and TUNEL method were used to observe the growth and apoptosis of the cells, respectively. Electrophoretic mobility shift assay （EMSA） and Western blot were performed to detect the activation of NF-kB and its inhibi- tion after Ad-IkBαM transfection, respectively. RESULTS： MTT indicated that the proliferations of BEL-7402 and SMMC-7721 cells were significantly suppressed after treatment of arsenious acid with different concentrations （P 〈 0.05）. Western blot and EMSA showed that arsenious acid markedly inhibited the growth of liver cancer cells, and promoted the activation of NF-kB. After transfection with Ad-IkBαM, the activation of NF-kB induced by arsenious acid was dramatically inhibited. The apoptosis rates were 66.47% and 36.67% in SMMC-7721 cells transfected with Ad-IkBαM and Ad-IkBα, respectively, and they were 74.5% and 32.37% in Bel-7402 cells transfected with Ad-IkBαM and Ad-IkBα, respectively. CONCLUSION： Arsenious acid has obvious effect in the treatment of hepatocarcinoma, but it promotes the activation of NF-kB at the same time. Recombinant adenovirus Ad-IkBαM can inhibit NF-kB activation while increase the effect of Arsenious acid effectively.

关 键 词：核转录因子KB 肝癌 亚砷酸 基因治疗 重组腺病毒 

分 类 号：R735.7[医药卫生—肿瘤]