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Killing Effects of Different Prodrugs Combined With Suicide Gene HSV-tk on Hep-2 in Vitro
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- Author:
- No author available
- Journal Title:
- JOURNAL OF CHINESE PHYSICIAN
- Issue:
- 3
- DOI:
- 10.3760/cma.j.issn.1008-1372.2003.03.002
- Key Word:
- 喉癌;HSV-tk基因;基因治疗
Abstract： 目的观察并比较不同前药与自杀基因HSV-tk共同作用对喉癌细胞Hep-2的体外杀伤作用.方法构建tk基因逆转录表达载体pL(tk)SN,在LipofectAMINETM2000介导下转染PA317包装细胞,G418筛选,直至出现抗性克隆,扩大培养,用NIH3T3测定病毒滴度,将重组病毒分别感染人喉癌细胞Hep-2,G418筛选出抗性克隆命名为Hep/tk,以RT-PCR及Southern blot检测tk基因的整合及表达.通过观察tk基因修饰细胞生长状况及比较不同前药对喉癌细胞杀伤效应以探讨不同前药作用机理.结果 RT-PCR及Southern blot分析证明tk基因在人喉癌细胞系Hep-2中有整合及表达;Hep/tk 与未转基因的原肿瘤细胞相比,二者生长速度及形态结构无明显差别;在不同前药敏感性试验中,Hep/tk显示出对GCV的高敏感性,而tk阳性和tk阴性细胞均显示对ACV和BVdU相对不敏感,tk基因修饰细胞与不同比例亲本细胞混合后经GCV治疗均显示出明显旁观者效应.结论人喉癌细胞系Hep-2对HSV-tk/GCV系统高度敏感,并且有明显的旁观者效应,可望成为喉癌基因治疗的新途径.
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