Construction of eukaryotic expression vector of mouse IFN-? and its antitumor effect
- VernacularTitle:小鼠IFN-?真核表达载体的构建及其抗肿瘤效应研究
- Author:
Mingyao ZHAO
;
Kangdong LIU
;
Ziming DONG
;
Guoqiang ZHAO
;
Hongyan YANG
;
Youtian HUANG
;
Zhimin ZHENG
- Publication Type:Journal Article
- Keywords:
Gene transfection;
Expression vector;
Macrophage;
Interferon type Ⅱ
- From:
Chinese Journal of Pathophysiology
1986;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct a mouse IFN-? expression vector and observe the antitumor effects of mouse peritoneal macrophages transfected with IFN-? in vivo and in vitro. METHODS: The IFN-? mRNA was amplified by RT-PCR. The open reading frame of mouse IFN-? gene was recombinanted with eukaryotic expression vector pcDNA3.1 through subcloning. Mouse peritoneal macrophages were transfected with recombinant vector pcDNA3.1-IFN-?. The expression of INF-? mRNA was measured by RT-PCR. Another group of peritoneal macrophages were cultured with the culture medium from pcDNA3.1-IFN-? transfecting groups, and its antitumor effect was measured by MTT. pcDNA3.1-IFN-? plasmid was peritoneally injected inte mouse with tumor. The appearance of ascites of pcDNA3.1-IFN-? plasmid injected mice and survival time were observed. RESULTS: The mouse IFN-? expression vector pcDNA3.1-IFN-? was constructed. The sequence was demonstrated to be the same as on GenBank. The recombinant vector was introduced into mouse peritoneal macrophages. IFN-? mRNA was detected by RT-PCR. The supernatant from cultured macrophages transfected with pcDNA3.1-IFN-? plasmid stimulated the antitumor effects of the macrophages without transfection. The appearance of ascites in pcDNA3.1-IFN-? plasmid injected mouse was delayed and survival time was longer than that in other groups. CONCLUSION: We have successfully constructed the mouse IFN-? expression vector pcDNA3.1-IFN-?. Mouse peritoneal macrophages transfected with pcDNA3.1-IFN-? have antitumor effects in vivo and in vitro.
