腺病毒介导HSV-tk自杀基因联合野生型p53基因对直肠癌细胞的杀伤作用

Antitumor Effects of Adenovirus Mediated Coinfection of HSV tk Suicide Gene and Wild type p53 Gene on Rectal Cancer Cells

目的 观察重组腺病毒介导单纯疱疹病毒胸苷激酶 (HSV tk)基因联合人野生型p5 3基因共转染 ,对直肠癌细胞的杀伤作用。方法 构建重组腺病毒质粒 pAdCMV Link1(tk/ p5 3)、pAdCMV Link1(tk)、pAdCMV Link1(p5 3) ,分别感染p5 3突变的人直肠癌细胞SW 837。进行细胞集落形成实验、细胞存活率的测定和裸鼠移植瘤治疗实验 ,观察HSV tk/GCV系统与野生型p5 3基因联合对肿瘤细胞的杀伤作用。 结果 应用 pAdCMV Link1(tk/ p5 3)、pAdCMV Link1(-)、pAdCMV Link1(tk)、pAd CMV Link1(p5 3)重组腺病毒感染SW 837细胞 ,加入GCV ,各组细胞集落数分别为 8、95、40、70。pAdCMV Link1(CD / p5 3)组肿瘤细胞集落形成减少、细胞存活率显著下降 (P <0 .0 1)。裸鼠移植肿瘤生长抑制率分别为 76 .5 %、0 .8%、5 5 .8%、2 3 .2 % ,pAd CMV Link1(tk/ p5 3)重组腺病毒对肿瘤的抑制作用最强。 结论 HSV tk自杀基因与野生型 p5 3基因共转染 。

Objective To investigate the antitumor effects of coinfection of herpes simple virus thymidine kinase suicide(HSV tk)gene/ganciclovir and human wild type p53 gene with adenovirus mediation on rectal cancer cells.Methods Recombinant adenovirus plasmid pAdCMV Link1(tk/p53)、pAdCMV Link1(-)、pAdCMV Link1(tk) and pAdCMV Link1(p53) were construeted and SW837 rectal cancer cells,which had p53 gene mutations were infected by them.Plating efficiency and MTT method were used and experimental treatment of xenografts of SW837 cells in nude mice was performed to evaluate antitumor effects of HSV tk/p53 gene coinfection.Results SW837 tumor cells were infected with recombinant adenovirus of pAdCMV Link1(tk/p53),pAdCMV Link1(-), pAdCMV Link1(tk) and pAdCMV Link1(p53).Adding GCV in each group,plating efficiency was 8,95,40,70 respectively.Decreases of plating efficiency and survival rates of tumor cells in pAdCMV Link1(tk/p53) recombinant adenovirus infection group had significant differences( P <0.01).Suppression rates of tumor growth of xenografts in nude mice were 76.5%, 0.8% ,55.8%,23.2% in ilka group.A high powered antitumor effect of uniting HSV tk/GCV with wild type p53 was observed.Conclution Coinfection of HSV tk and wild type p53 genes is able to exert combined effects of suicide gene and antioncogene,and has more powerful antitumor actions.