Experimental studies on effects of EGFR gene suppression by RNA interference on enhancing radiosensitivity of ovarian cancer cells

LUO Qian; et al

doi:10.13406/j.cnki.cyxb.2010.05.038

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Experimental studies on effects of EGFR gene suppression by RNA interference on enhancing radiosensitivity of ovarian cancer cells

更新时间：2024-01-17

- Experimental studies on effects of EGFR gene suppression by RNA interference on enhancing radiosensitivity of ovarian cancer cells
- Journal of Chongqing Medical University Vol. 35, Issue 5, Pages: 670-673(2010)
- 作者机构：
  
  1. 重庆医科大学附属第一医院肿瘤科
  2. 重庆医科大学临床学院重庆市肿瘤研究所
  3. 第三军医大学大坪医院肿瘤科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.05.038
  CLC： R737.31
- Published：2010，
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[1]罗茜,吴永忠,林远洪,郭启帅,黄环.RNA干扰EGFR基因对卵巢癌细胞放疗敏感性影响的实验研究[J].重庆医科大学学报,2010,35(05):670-673.

LUO Qian, et al. Experimental studies on effects of EGFR gene suppression by RNA interference on enhancing radiosensitivity of ovarian cancer cells. [J]. Journal of Chongqing Medical University 35(5):670-673(2010)
[1]罗茜,吴永忠,林远洪,郭启帅,黄环.RNA干扰EGFR基因对卵巢癌细胞放疗敏感性影响的实验研究[J].重庆医科大学学报,2010,35(05):670-673. DOI： 10.13406/j.cnki.cyxb.2010.05.038.

LUO Qian, et al. Experimental studies on effects of EGFR gene suppression by RNA interference on enhancing radiosensitivity of ovarian cancer cells. [J]. Journal of Chongqing Medical University 35(5):670-673(2010) DOI： 10.13406/j.cnki.cyxb.2010.05.038.

摘要

目的:构建针对表皮生长因子受体（Epidermal growth factor receptor

EGFR）基因的小于扰RNA（Small interference RNA

siRNA）质粒

转染人卵巢癌SKOV3细胞株后

检测RNA干扰（RNA interference

RNAi）对EGFR基因表达及评估细胞对放疗敏感性的影响。方法:体外构建EGFR小发卡状RNA（Short hairpin RNA

shRNA）的表达质粒

脂质体法转染入SKOV3细胞及G418筛选阳性克隆。实验随机分为阳性RNA干扰组（A组）、阴性RNA干扰组（B组）、空白对照组（C组）。采用RT-PCR、Western Blot技术检测EGFR mRNA及其蛋白质的表达情况。分别给予不同放射剂量（0、2、4、6、8Gy）照射

MTT法绘制细胞存活曲线。结果:靶向EGFR的序列特异性shRNA可明显抑制EGFR基因的表达

EGFR mRNA和蛋白的抑制率分别为77.5%和76.1%;序列特异性shRNA-EGFR可明显提高SKOV3细胞对放疗的敏感性

差异有统计学意义（P<0.01）。结论:体外研究表明RNAi沉默EGFR基因可以提高卵巢癌SKOV3细胞对放疗的敏感性。

Abstract

Objective:To construct a expression vector of EGFR specific siRNA and transfect it into human ovarian cancer cell line SKOV3

then to evaluate the effect of RNA interference(RNAi) on the target gene expression and radiosensitivity of the cell line.Methods:EGFR sh-RNA was synthesized in vitro and transfected into SKOV3 cells with lipofectamine technique and then the positive clones were selected by G418.The positive RNA interference cell line

negative RNA interference cell 1ine and the normal control cell 1ine were included in the study.RT-PCR and Western Blot technique were performed to detect the inhibitory effect of RNAi on EGFR mRNA level and protein level

respectively.Three kinds of cell lines were exposed to different doses of radiation.Cell survival curve was drawn by means of MTT method.Results:The expression of EGFR gene was surpressed obviously in the specific sequence shRNA group

and EGFR mRNA and protein surpression rates were 77.5% and 76.1%

respectively;Sequence-specific shRNA-EGFR can significantly enhance the radiosensitivity of ovarian cancer cells

and significant statistical differences existed among the three groups(P<0.01).Conclusion:The experiments in vitro confirmed that EGFR gene silenced by RNAi can significantly enhance the radiosensitivity of ovarian cancer SKOV3 cells.

关键词

EGFR基因RNA干扰卵巢癌放疗敏感性

Keywords

EGFR geneRNA interferenceovarian cancerRadiosensitivity

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