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

杨巍; 李艳博; 赵敬国; 龚守良; 曹建平; 刘芬菊

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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

研究论文 | 更新时间：2023-03-30

- 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
- 辐射研究与辐射工艺学报 2009年27卷第2期页码：120-124
- 作者机构：
  
  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
- 作者简介：
  
  杨巍，男，1976年10月出生，2005年毕业于吉林大学放射医学专业获博士学位，讲师，主要从事肿瘤的基因-放射治疗研究
- 基金信息：
  
  国家自然科学基金青年项目(30600160)
- DOI：
  中图分类号：
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杨巍, 李艳博, 赵敬国, 等. 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[J]. 辐射研究与辐射工艺学报, 2009,27(2):120-124.

YANG Wei, LI Yanbo, ZHAO Jingguo, et al. anti-tumor effect of silencing Hif-1α and Survivin genes combined with X-ray irradiation on hypoxic tumor cells in vitro[J]. Journal of Radiation Research and Radiation Processing, 2009,27(2):120-124.
杨巍, 李艳博, 赵敬国, 等. 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[J]. 辐射研究与辐射工艺学报, 2009,27(2):120-124. DOI：

YANG Wei, LI Yanbo, ZHAO Jingguo, et al. anti-tumor effect of silencing Hif-1α and Survivin genes combined with X-ray irradiation on hypoxic tumor cells in vitro[J]. Journal of Radiation Research and Radiation Processing, 2009,27(2):120-124. DOI：

摘要

为探讨RNA干扰（RNA interference，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36 h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。

Abstract

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 cells 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 transfection 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

Keywords

RNA interferenceX-rayHIF-1αSurvivin

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