siRNA沉默肾癌细胞EGFR基因对放疗敏感性影响的研究

褚晗; 于德新; 张志强; 黄韵; 谢栋栋; 王毅; 张涛; 陈磊; 丁德茂; 邹慈; 闵捷; 王大明

siRNA沉默肾癌细胞EGFR基因对放疗敏感性影响的研究

- 1. 安徽医科大学第二附属医院泌尿外科(合肥, 230601);
- 2. 合肥微尺度物质科学国家实验室中国科技大学生命科学院分子细胞遗传学实验室
基金项目:
安徽省高校省级自然科学研究项目（编号ZD200907）

详细信息

通讯作者: 于德新,E-mail:yudx_urology@yahoo.com

中图分类号: R737.11

收稿日期: 2012-08-14

The effects of EGFR gene RNAi on the radiosensitivity of renal cell carcinoma

- 1. Department of Urology, Second Affiliated Hospital of Anhui Medical University, Hefei, 230601, China;
- 2. Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China

More Information

Corresponding author: YU Dexin, E-mail: yudx_urology@yahoo.com

Received Date: 14 August 2012

摘要

摘要: 目的：探讨siRNA沉默ACHN肾癌细胞的表皮生长因子受体（epidermal growth factor receptor， EGFR）对放疗敏感性的影响。方法：免疫组化及细胞免疫荧光技术证明组织水平及细胞水平EGFR的表达，化学合成针对EGFR的小干扰RNA，通过脂质体转染法转染进ACHN肾癌细胞中，利用Western blot技术检测细胞中EGFR蛋白的表达，然后分别用0、2、4、6、8Gy剂量的X线对5组肾癌细胞（每组包含空白对照组、非异性RNA干扰组、特异性RNA干扰组）进行照射，光学显微镜下观察细胞凋亡情况，采用台盼蓝据染法检测细胞凋亡率。结果：靶向EGFR序列的特异性干扰RNA可明显抑制EGFR蛋白的表达，光学显微镜下观察到RNAi联合放疗组细胞死亡情况明显，台盼蓝拒染法检测特异性干扰EGFR基因组可显著提高ACHN肾癌细胞对放疗的敏感性（P<0.05）。结论：体外研究表明，siRNA沉默ACHN肾癌细胞的EGFR可明显提高肾癌细胞对放疗的敏感性，为肾癌放射治疗提供了新思路。
- 肾癌 /
- 表皮生长因子受体(EGFR) /
- RNAi /
- 放疗
Abstract: Objective: To study the effects of down regulation of epidermal growth factor receptor(EGFR)gene on the radiosensitivity of renal cell carcinoma.Method: We used immunohistochenmical and immunofluorescence to examine the level of EGFR expression in renal cell carcinoma. ACHN cells were transfected with small interfering RNA (siRNA) by Liposome transfection reagent targeting human EGFR. The expression level of EGFR was detected by Western blot analysis. The ACHN cells are divided into two groups, 1):single radiotherapy and 2):RNAi combined with radiotherapy.Both of the groups were irradiated with 0, 2, 4, 6, 8 Gy X-ray. We observed cell morphology under opti-microscope. The cell death curves were determined by trypan blue staining.Result: Sequence-specific shRNA targeting EGFR gene could suppress the expression of EGFR gene remarkably; Cell death were more prominent in group 2) than in group 1).Conclusion: EGFR RNAi significantly enhanced radiosensitivity of renal carcinoma cells.
- renal cell carcnimoma /
- EGFR gene /
- RNA interference /
- radiotherapy

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