The inhibition of intrinsic expression of GFP in nasopharyngeal carcinoma by RNA interferencing
- VernacularTitle:RNA干扰技术抑制鼻咽癌细胞内源性绿色荧光蛋白的表达
- Author:
Yiqin WANG
- Publication Type:Journal Article
- Keywords:
Nasopharyngeal neoplasms;
Green fluorescent protein;
Short hairpin RNA;
RNA interference
- From:
Journal of Chongqing Medical University
2007;0(08):-
- CountryChina
- Language:Chinese
-
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.
