Construction of retroviral vectors to induce strong hepatoma cell-specific expression of murine tumor necrosis factor gene and the hepatoma-specifie gene therapy mediated by the recombinant retroviral vectors.
- VernacularTitle:肿瘤坏死因子肝癌特异性重组载体的构建及其介导的肝癌特异性基因治疗的研究
- Author:
Guangwen CAO
;
Ping DU
;
Wenguo YANG
- Publication Type:Journal Article
- Keywords:
Hepatoma Murine tumor necrosis factor Hepatoma-specific transcriptional regulatory sequence Retroviral vectors Murine Gene therapy
- From:
Chinese Journal of Digestion
1996;0(S1):-
- CountryChina
- Language:Chinese
-
Abstract:
Murine tumor necrosis factor(mTNF)cDNA was inserted into the polylinker site of MNSM retroviral vector to create pMNSM-SV40-mTNF. Albumin enhancer/promoter (Alb e/p) sequence was used to replace SV40 early region promoter of pMNSM-SV40-mTNF vector to create pMNSM-Alb e/p-mTNF recombinant retroviral vector. The retroviral constructs were introduced into amphotropic retroviral packaging cells PA317. Production of the recombinant retroviruses was accomplished by the lipofectamine-mediated gene transfer procedure. The murine tumor cells were infected with the retroviruses in the presence of polybrene. Dot hybridization of total RNA from modified tumor cell with mTNF cDNA probe and mTNF bioassay demonstrated that transcription and expression of mTNF gene drived by Alb e/p were markedly high in the hepatoma cells which produced albumin, and inhibited in the non-hepatoma tumor cells. The hepatoma cells modified with mTNF gene lost its tumorgenicity and significantly inhibited the growth of the parental hepatoma in vivo. High-titer MNSM-Alb e/p-mTNF retroviruses or the high-titer retroviruses producing packaging cells, after intra-tumoral injection, specifcally inhibited the growth of the hepatoma, and significantly prolonged the survival period of the hepatoma-loaded mice. Biopsy and immunohistochemical assay of the hepatoma during in vivo gene therapy, showed the occurrence of extensive tumor necrosis, bleeding, and CD8+, CD25+, CD4+ lymphocytic infiltration and fibrosis.
