建立稳定表达融合自杀基因CD/UPRT.UL49的CNE-2鼻咽癌细胞株

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收稿日期: 2013-03-26

Construction of nasopharyngeal carcinoma CNE-2 cell lines expressing stable fusion suicide gene CD/UPRT.UL49

- Department of Otolaryngology, Xiangya Hospital of Central South University, Changsha, 410008, China

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Received Date: 26 March 2013

摘要: 目的:建立稳定表达融合自杀基因CD/UPRT.UL49的CNE-2鼻咽癌细胞株。方法:构建表达载体pcDNA3.1(-)E6.BARF1p.CD/UPRT.UL49,经脂质体法转染CNE-2细胞,G418和前体药物5-氟胞嘧啶筛选出阳性克隆细胞并扩大培养。抽提阳性克隆细胞总蛋白质,Western-blotting检测目的基因表达。结果:融合自杀基因CD/UPRT.UL49在CNE-2转染细胞内稳定表达。结论:本组通过脂质体转染技术,成功地建立了稳定表达融合CD/UPRT.UL49基因的CNE-2细胞株。
- 自杀基因 /
- CD/UPRT.UL49 /
- CNE-2细胞 /
- 鼻咽肿瘤
Abstract: Objective:To construct nasopharyngeal carcinoma CNE-2 cell lines expressing stable fusion suicide gene CD/UPRT.UL49.Method:The plasmids of pcDNA3.1(-)E6.BARF1p.CD/UPRT.UL49 was transfected into CNE-2 cells through lipofectamine, and the transfected CNE-2 cells were selected by G418 and prodrugs for getting the cells expressing fusion CD/UPRT.UL49 gene. The protein produced by the suicide gene was tested by Western-blotting in CNE-2 cells.Result:Suicide genes were expressed stably in CNE-2 cells.Conclusion:We constructed nasopharyngeal carcinoma cell lines CNE-2 expressing stable suicide gene through lipofectamin.
- suicide gene /
- CD/UPRT.UL49 /
- CNE-2cells /
- nasopharyngeal neoplasm

[1]	SPANO J P, BUSSON P, ATLAN D, et al.Nasopharyngeal carcinomas:an update[J].Eur J Cancer, 2003, 39:2121-2135.
[2]	PAK M W, TO K F, LO Y M, et al.Nasopharyngeal carcinoma in situ (NPCIS) -pathologic and clinical perspectives[J].Head Neck, 2002, 24:989-995.
[3]	SANCHEZ-PEREZ L, KOTTKE T, DANIELS G A, et al.Killing of normal melanocytes, combined with heat shock protein 70and CD40 Lexpression, cures large established melanomas[J].J Immunol, 2006, 177:4168-4177.
[4]	BOSENBERG M, MUTHUSAMY V, CURLEY D P, et al.Characterization of melanocyte-specific inducible Cre recombinase transgenic mice[J].Genesis, 2006, 44:262-267.
[5]	卿菁, 赵素萍, 蒋卫红, 等.BARF1基因的启动子活性及其在鼻咽癌细胞中肿瘤特异性分析研究[J].现代实用医学, 2013, 25 (5):492-494.
[6]	唐瑶云, 肖健云, 赵素萍.靶向性质粒表达载体pcDNA3.1 (-) CMV Egr-1.CDglyTK的构建及转染研究[J].中国耳鼻咽喉颅底外科杂志, 2003, 9 (1):34-37.
[7]	BOENICKE L, CHU K, PAULS R, et al.Efficient dose-dependent and time-dependent protein transduction of pancreatic carcinoma cells in vitro and in vivo using purified VP22-EGFP fusion protein[J].J Mol Med (Berl), 2003, 81:205-213.
[8]	XIONG F, XIAO S, YU M, et al.Enhanced effect of microdystrophin gene transfection by HSV-VP22 mediated intercellular protein transport[J].BMC Neurosci, 2007, 8:50-50.