含HSV1-TK基因重组腺病毒的构建及对小鼠肺癌的抑制作用

Construction of recombinant adenovirus vector with HSV1-TK gene and the inhibitory effect on mice lung carcinoma

目的 观察含单纯疱疹病毒 1型胸苷激酶 (HSV1- TK)基因和 CMV启动子的重组腺病毒Ad E1 CMVHSV1- TK(Ad TK)对 L795小鼠肺癌细胞和 T739近交纯系小鼠肺癌的抑制作用。方法 采用同源重组的方法在人胚肾 2 93细胞中构建重组腺病毒 Ad TK ,用 PCR方法鉴定 ;TCID50 (5 0 %组织细胞感染量 )方法滴定病毒 ;将荷瘤小鼠分为 DMEM对照组 ,Ad L ac Z对照组 ,Ad TK实验一组 ,Ad TK实验二组 ;连续观察各组小鼠肿瘤的生长速度以及存活期。结果 PCR证实了 TK基因的导入 ;动物实验结果显示实验组小鼠瘤体的生长速度低于对照组 ,且生存期长于对照组 ,均有统计学差异。肿瘤组织病理切片显示对照组肿瘤组织比实验组生长旺盛。结论 可在人胚肾 2 93细胞中同源重组构建重组腺病毒 Ad TK,且能达到治疗有效滴度 ;Ad TK/ GCV系统对小鼠肺癌具有明显的治疗效果。

Objective To construct recombinant replication-defective adenovirus (AdTK) containing HSV1-TK gene and to study on its inhibitory effect on mice lung carcinoma. Methods AdTK was constructed using homologous recombinant method in human embryo kidney 293 cells,identified by PCR and titered by TCID 50 assay.Mice tumor model was established by transplanting LA795 mice lung cancer cells into T739 inbred mice.The mice bearing tumors were randomized into four groups:DMEM control group,AdlacZ control group,AdTK treating group 1,and AdTK treating group 2.Survival and tumor growth rate were observed for each group. Results AdTK successfully constructed reached significant titer.It was demonstrated that there were AdTK/GCV treatment suppresses in tumor growth rate and prolongation of survival period compared with the two control groups (P<0.01). Conclusion Recombinant adenovirus can be constructed using homologous recombinant method in human embryo kidney 293 cells, and can reach significant titer.AdTK/GCV system has significant inhibitory effect on mice lung carcinoma.