The inhibition of intrinsic expression of GFP in nasopharyngeal carcinoma by RNA interferencing

WANG Yiqin; et al

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The inhibition of intrinsic expression of GFP in nasopharyngeal carcinoma by RNA interferencing

更新时间：2024-01-17

- The inhibition of intrinsic expression of GFP in nasopharyngeal carcinoma by RNA interferencing
- Journal of Chongqing Medical University Issue 8, Pages: 822-825(2007)
- 作者机构：
  
  1. 重庆医科大学附属第一医院耳鼻咽喉头颈外科
  2. 重庆医科大学附属第一医院眼科
- 作者简介：
- 基金信息：
- DOI：
  CLC： R739.63
- Published：2007，
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[1]王祎琴,彭惠,洪苏玲.RNA干扰技术抑制鼻咽癌细胞内源性绿色荧光蛋白的表达[J].重庆医科大学学报,2007(08):822-825+834.

WANG Yiqin, et al. The inhibition of intrinsic expression of GFP in nasopharyngeal carcinoma by RNA interferencing. [J]. Journal of Chongqing Medical University (8):822-825(2007)
[1]王祎琴,彭惠,洪苏玲.RNA干扰技术抑制鼻咽癌细胞内源性绿色荧光蛋白的表达[J].重庆医科大学学报,2007(08):822-825+834. DOI：

WANG Yiqin, et al. The inhibition of intrinsic expression of GFP in nasopharyngeal carcinoma by RNA interferencing. [J]. Journal of Chongqing Medical University (8):822-825(2007) DOI：

摘要

目的:建立稳定表达绿色荧光蛋白（GFP）的鼻咽癌细胞株;观察不同转染浓度的短发卡状RNA对鼻咽癌细胞内源性GFP的抑制效率及细胞毒性。方法:转染pEGFP-N1至鼻咽癌细胞株HNE1

G418筛选获得稳定表达GFP的HNE1-GFP细胞株;利用针对GFP基因的短发卡状RNA表达载体shGFP

以不同浓度shGFP转染HNE1-GFP

荧光显微镜观察细胞荧光强度

软件分析荧光抑制效率

MTT检测细胞毒性

RT-PCR检测GFPmRNA水平改变

Western Blot检测GFP蛋白水平改变。结果:绿色荧光稳定表达于传代HNE1细胞中;shGFP能够显著抑制该细胞中GFP的表达

抑制效率分别为49.7%

84.8%

86.7%且无细胞毒性;GFP在mRNA和蛋白水平的表达均有明显下降。结论:稳定表达GFP的鼻咽癌细胞株成功建立;RNA干扰载体可抑制鼻咽癌细胞中GFP的表达

其抑制效率有明显的浓度依赖性且无明显细胞毒性。该系统能够用于鼻咽肿瘤的RNA干扰机制研究中。

Abstract

Objective:To construct a nasopharyngeal carcinoma cell strain expressing GFP stably and observe the inhibition effect and cytotoxicity of short hairpin RNA targeting the intrinsic GFP.Methods:HNE1 cells transfected with pEGFP-N1 were followed by the screening with G418 to obtain the stable nasopharyngeal carcinoma cell strain expressing GFP(HNE1-GFP);shGFP

a vector expressing short hairpin RNA targeting GFP

was transfected into HNE1-GFP with various levels.The fluorescence was observed by fluorescent microscope.The inhibition ratios and cytotoxicity of shRNA were analyzed by software and MTT

respectively;The protein and mRNA level of GFP were detected by Western Blot and RT-PCR

respectively.Results:The GFP gene was expressed stably within HNE1-GFP cell line;shGFP suppressed the intrinsic expression of GFP with the ratios of 49.7%

84.8% and 86.7%

respectively

without significant cytotoxicity.Conclusion:A nasopharyngeal carcinoma cell strain expressing GFP stably is established successfully;The inhibition effect of RNA interference is demonstrated in nasopharyngeal carcinoma cells and it is characterized by the significant concentration dependence and rare cytotoxicity.

关键词

鼻咽肿瘤绿色荧光蛋白短发卡状RNARNA干扰

Keywords

Nasopharyngeal neoplasmsgreen fluorescent proteinShort hairpin RNARNA interference

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