Construction of recombinant adenovirus vector with HSV-TK controlled by hTERT promoter and experimental study of anti-hepatoma
10.3760/cma.j.issn.1006-9801.2009.01.002
- VernacularTitle:重组腺病毒Ad-hTERTp-HSV-TK载体的构建及抗肝癌实验研究
- Author:
Yan LIU
;
Zhihua DENG
;
Chongqing YANG
;
Jinglong LIU
;
Ring JIA
;
Suyo GUO
;
Qiang YANG
- Publication Type:Journal Article
- Keywords:
hTERT promoter;
Simplexvirus;
Thymidine kinase;
Adenovirus vector;
Liver neoplasms;
Gone;
Therapy
- From:
Cancer Research and Clinic
2009;21(1):4-6
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the recombinant adenovirus vector with hTERT-HSV-TK and observe the killing effect of Ad-hTERTp-HSV-TK/GCV system on hepatocellular carcinoma cells. Methods A recombinant replication defective adenoviral vector of Ad-hTERTp-HSV-TK was constructed via homologous recombination which both shuttle plasmid pSU-Tp-TK and adenovirus backbone plasmid pBHGE3 transfected into the HEK293 packaging cells. Then the Ad-hTERTp-HSV-TK was amplified and purified through PCR. The activity of the HepG2 cells and the L-02 cells were tested by methyl thiazolyl terazolium (MTT) after they were transfected by the recombinant adenovirus of different multiplicities of infection (MOI) and then were added GCV of different conc.entration. Results The recombinant replication defective adenoviral vector of Ad-hTERTp-HSV-TK were identified by PCR successfully. The viral titer was 1.5×1010 pfu/ml after amplification and purification. The HepG2 cells were targetedly suppressed by Ad-hTERTp-HSV-TK/GCV system. The survival rate of cells decreased gradually along with the increase of the MOI and the GCV' s concentration. Conclusion The recombinant replication defective adenoviral vector of Ad-hTERTp-HSV-TK can inhibit the HepG2 cells significantly, but has not influence on the L-02 cells.
