沉默乏氧诱导因子-1α和生存素基因联合X射线照射对乏氧细胞的体外抑瘤效应  

Anti-tumor effect of silencing Hif-1α and Survivin genes combined with X-ray irradiation on hypoxic tumor cells in vitro

作　　者：杨巍[1] 李艳博[2] 赵敬国[3] 龚守良[2] 曹建平[1] 刘芬菊[1] 

机构地区：[1]苏州大学放射医学与公共卫生学院放射生物学教研室,苏州215123 [2]吉林大学公共卫生学院卫生部放射生物学重点实验室,长春130021 [3]中国人民解放军403医院整形外科,大连116015

出　　处：《辐射研究与辐射工艺学报》2009年第2期120-124,共5页Journal of Radiation Research and Radiation Processing

基　　金：国家自然科学基金青年项目(30600160)资助

摘　　要：为探讨RNA干扰(RNAinterference,RNAi)沉默乏氧诱导因子-1α(Hypoxia induced factor 1α,HIF-1α)和生存素(Survivin)基因联合X射线照射对乏氧人肝癌细胞SMMC-7721的体外抑瘤效应,利用双靶向HIF-1α和Survivin基因载体,采用阳离子脂质体介导法转染SMMC-7721细胞,经乏氧培养36h后,分别以RT-PCR和Western blot法检测HIF-1α和Survivin基因的mRNA及其蛋白表达,分别以四甲基偶氮唑盐(MTT)法和流式细胞术检测细胞存活分数及细胞凋亡。结果表明,转染质粒pGenesil-Survivin-HIF的SMMC-7721细胞中,HIF-1α和Survivin基因mRNA表达水平与对照和阴性干扰组相比明显降低(p<0.05),也未检测到HIF-1α和Survivin蛋白的表达。单、双干扰联合放疗组存活分数较照射组明显降低(p<0.01),双干扰联合放疗组存活分数较其它组也明显降低(p<0.01)。单、双干扰联合放疗组凋亡细胞百分数较对照组明显升高(p<0.01),双干扰联合放疗组凋亡细胞百分数较其它组也明显升高(p<0.01)。结果提示,质粒pGenesil-Survivin-HIF可有效地干扰乏氧SMMC-7721细胞HIF-1α和Survivin基因的mRNA及其蛋白表达。双干扰联合放疗组对乏氧SMMC-7721细胞的体外抑瘤效应明显优于单干扰联合放疗组。In order to investigate the anti-tumor effect of RNA interference silencing HIF- 1α and Survivin genes combined with X-ray irradiation on hypoxic SMMC-7721 cells in vitro, we detected mRNA and protein expression of HIF-1α and Survivin genes in hypoxic SMMC-7721 ceils after transfection with pGenesil-Survivin-HIF carried by liposome by RT-PCR and Western blot respectively. Survival fraction and apoptosis of hypoxic SMMC-7721 cells in vitro were detected by MTT and FCM assay respectively. The results show that mRNA expression of HIF-1α and Survivin genes in hypoxic SMMC-7721 cells transfected with pGenesil-Survivin-HIF is significantly lower than that of control and negative interference group （p〈0.05）. Protein expression of HIF-1α and Survivin genes in hypoxic SMMC-7721 cells after transfectlon with pGenesil-Survivin-HIF can not be detected. Survival fraction of pGenesil-Survivin＋5 Gy group, pGenesil-HIF＋5 Gy group and pGenesil-Survivin-HIF＋5 Gy group is significantly lower than that of 5 Gy group （p〈0.01）. Survival fraction of pGenesil-Survivin-HIF＋5 Gy group is also significantly lower than that of other groups （p〈0.01）. Percentage of apoptosis of pGenesil-Survivin＋5 Gy group, pGenesil-HIF＋5 Gy group and pGenesil-Survivin-HIF＋5 Gy group is significantly higher than that of control group （p〈0.01）. Percentage of apoptosis fraction of pGenesil-Survivin-HIF＋5 Gy group is significantly higher than that of other groups （p〈0.01）. These results can be suggested that the pGenesil-Survivin-HIF could silence mRNA and protein expression of HIF-1α and Survivin genes in hypoxic SMMC-7721 cells. Anti-tumor effect of pGenesil-Survivin-HIF＋5 Gy group is more powerful than that of pGenesil-Survivin＋5 Gy group or pGenesil-HIF＋5 Gy group with hypoxic SMMC-7721 cells in vitro.

关 键 词：RNA干扰 X射线 乏氧诱导因子-1Α 生存素 

分 类 号：R730.54[医药卫生—肿瘤] R730.55[医药卫生—临床医学]