摘要
目的 研究HSV tk/GCV系统对NIH3T3细胞系的杀伤作用 ,为与成纤维细胞增殖有关的皮肤病基因治疗提供体外实验依据。方法 应用携带HSV tk基因的重组质粒pPNT tk通过磷酸钙转染传代培养的NIH3T3细胞 ,并经G418筛选 ,获得抗药性细胞集落 ,利用MTT法检测不同浓度GCV对NIH 3T3细胞的杀伤作用。结果 GCV对NIH3T3 /tk细胞有明显的细胞毒性作用 ,当GCV浓度为 10 .0 μmol/L时 ,NIN3T3 /tk细胞的生存抑制率已高达 95 %以上。而在同样药物浓度条件下 ,未转染的对照NIH 3T3细胞的生存抑制率为 11.9% ,与空白对照组相比较 ,差异无显著性。结论 HSV tk/GCV系统对NIH3T3细胞有很好的抑制作用 ,转染后细胞较之未转染细胞对前体药物更昔洛韦的敏感度显著增加 ,且呈剂量依赖性。
Objective To investigate the cytotoxic effect of HSV tk/GCV system of NIH 3T3 cell line and to provide in vitro experimental evidence to the gene therapy of diseases related to fibroblast proliforation. Methods The recombinant plasmid containing HSV tk gene (pPNT tk) was transfected into NIH 3T3 cell by Ca3(PO4)2.Then,the cytotoxic efficacy of HSV tk/GCV system of 3T3 cell line was evaluated by MTT method.Results The recombinant plasmid pPNT tk can transfer HSV tk gene into 3T3 cell line and make it sensitive to GCV.The growth inhibitory rate of 3T3 cell line transfected by pPNT tk was 95.1% in the medium containing 10 μmol/L GCV,while that of the control 3T3 cells was 11.9%.Conclusion MTT assays showed that after HSV tk transfection,the susceptibility of NIH 3T3 cells to GCV was much higher than prior to transfection.Morever,the cytotoxic effect on 3T3/tk cell proliferation by GCV was dose dependent.
出处
《中国皮肤性病学杂志》
CAS
北大核心
2002年第6期368-369,373,共3页
The Chinese Journal of Dermatovenereology
基金
西安交通大学"教育振兴行动计划"重点培植项目基金资助 (PZ0 4 5)
