Killing Effects of Different Prodrugs Combined With Suicide Gene HSV-tk on Hep-2 in Vitro
- VernacularTitle:不同前药联合HSV-tk自杀基因系统对喉癌细胞体外杀伤效应的比较研究
- Author:
Lu YAN
;
Yan LI
;
Min SHEN
- Publication Type:Journal Article
- Keywords:
Laryngocarcinoma;
HSV-tk gene;
Gene therapy
- From:
Journal of Chinese Physician
2001;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate and compare the killing effects of different prodrugs combined with suicide gene HSV-tk on laryngocacinoma cell, Hep-2 in vitro. Methods Retroviral expressing vector pL(tk)SN was constructed by recombinant DNA technology. Hep-2 cells were infected by the recombinant retrovirus. The positive cloning was obtained after G418 selection and were termed Hep/tk. The integration and expression of tk gene in Hep-2 cells were identified by RT-PCR and Southern blot. The growth state and prodrugs killing effect of tk gene modified cells were used to investigate the expression of tk gene and antitumour effect on Hep-2 cells. Results RT-PCR and Southern blot analysis confirmed the integration and expression of tk gene in Hep-2 cells. There was no significant difference in cell proliferation between the Hep/tk and Hep-2. After the treatment of GCV,the Hep/tk showed high sensitivity to GCV and bystander effects were observed siginificantly in vitro. However the efficiency of another two prodrugs ACV and BVdU was lower than that of GCV. Both tk-positive and tk-negative Hep-2 cells were relatively insensitive to ACV and BVdU.Treatment of tk gene modified cells mixed with different proportion parental cells shoewd obviously bystander effects.Conclusions The laryngocarcinoma cells Hep-2 have sensitive to HSV-tk/GCV system and have significant bystander effects, which might have therapeutic potential value for laryngocarcinoma.
