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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2016, Vol. 36 Issue (2): 16-21    DOI: 10.13523/j.cb.20160203
研究报告     
PBI-SUR-TK载体靶向介导HSV-TK自杀基因诱导肝癌细胞凋亡
张英敏1, 赵娜1, 李勇芳1, 孟凡秀1, 张琪1, 高然朋1, 张悦红1, 于保锋1, 郭睿1, 王海龙1, 解军1, 徐钧2
1. 山西医科大学基础医学院 太原 030001;
2. 山西医科大学附属山西省肿瘤医院 太原 030013
Targeted Treatment of Hepatoma Using HSV-TK Suicide Gene with The PBI-SUR-TK Vector
ZHANG Ying-min1, ZHAO Na1, LI Yong-fang1, MENG Fan-xiu1, ZHANG Qi1, GAO Ran-peng1, ZHANG Yue-hong1, YU Bao-feng1, GUO Rui1, WANG Hai-long1, XIE Jun1, XU Jun2
1. Biochemistry and Molecular Biology, Shanxi Medical University, Taiyuan 030001, China;
2. Affiliated Tumor Hospital of Shanxi Medical University, Taiyuan 030013, China
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摘要:

目的:构建Survivin启动子调控的表达载体,并检测在启动子调控下HSV-TK自杀基因对肝癌细胞HepG2和正常肝细胞HL-7702凋亡的影响。方法:合成含TK基因的质粒PBI-SUR-TK,利用脂质体Lipofectamine 2000将其导入肝癌细胞和肝细胞。然后分别运用RT-PCR和Western blot特异性检测基因和蛋白的表达情况;利用CCK8方法检测细胞增殖情况,流式细胞仪上机检测细胞凋亡情况。结果:肝癌细胞转染组有更多的TK基因表达产物,增殖情况减弱,凋亡情况明显。结论:Survivin启动子驱动的HSV-TK/GCV自杀基因系统对肝癌可能有一定的治疗作用。

关键词: 肝癌凋亡Survivin启动子HSV-TK/GCV    
Abstract:

Objective:To construct a kind of expression vector which is cotrolled by survivin promoter, making a survivin promotor driving HSV-TK/GCV suicide gene system, and then detect its apoptosis inducing effect on HepG2 and HL-7702 cells. Methods:The study group synthesized a plasmid PBI-SUR-TK which contains TK gene, and transfect it into HepG2 and HL-7702 cells by liposome Lipofectanfine 2000.The HSV-TK gene expression by RT-PCR and the proteinlevel by Western blotting were detected. After adding GCV, CCK8 and flow cytometry were used to analyse the cell proliferation and apoptosis. Results:Contrasted with the control group, the HepG2 experimental group expressed more TK gene products, less proliferation and more apoptosis. There was no obvious difference between the HL-7702 groups. Conclusion:The HSV-TK/GCV suicide gene system driven by survivin promoter possibly has therapeutical effects on hepatoma.

Key words: HSV-TK/GCV    Hepatoma    Survivin promoter    Apoptosis
收稿日期: 2015-08-13 出版日期: 2015-11-19
ZTFLH:  Q789  
基金资助:

国家自然科学基金(30901821,81172136)、山西省自然科学基金(2015011113)资助项目

通讯作者: 于保锋, 徐钧     E-mail: shanxiyangcheng@126.com;junxuty@163.com
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王海龙
解军
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引用本文:

张英敏, 赵娜, 李勇芳, 孟凡秀, 张琪, 高然朋, 张悦红, 于保锋, 郭睿, 王海龙, 解军, 徐钧. PBI-SUR-TK载体靶向介导HSV-TK自杀基因诱导肝癌细胞凋亡[J]. 中国生物工程杂志, 2016, 36(2): 16-21.

ZHANG Ying-min, ZHAO Na, LI Yong-fang, MENG Fan-xiu, ZHANG Qi, GAO Ran-peng, ZHANG Yue-hong, YU Bao-feng, GUO Rui, WANG Hai-long, XIE Jun, XU Jun. Targeted Treatment of Hepatoma Using HSV-TK Suicide Gene with The PBI-SUR-TK Vector. China Biotechnology, 2016, 36(2): 16-21.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20160203        https://manu60.magtech.com.cn/biotech/CN/Y2016/V36/I2/16

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