Construction of adenovirus vector expressing IL-24 and E1A and its inhibition of SMMC-7721.
- Author:
Xiaohua WANG
1
;
Jingcheng MIAO
;
Yufeng XIE
;
Weihua SHENG
;
Yunbo SHAN
;
Jicheng YANG
Author Information
1. No. 1 Affiliated Hospital of Soochow University, Suzhou 215001, China.
- Publication Type:Journal Article
- MeSH:
Adenoviridae;
genetics;
metabolism;
Adenovirus E1A Proteins;
biosynthesis;
genetics;
Antineoplastic Agents;
pharmacology;
Apoptosis;
drug effects;
Carcinoma, Hepatocellular;
pathology;
Cell Line, Tumor;
Cell Proliferation;
Genetic Vectors;
genetics;
Humans;
Interleukins;
biosynthesis;
genetics;
Liver Neoplasms;
pathology;
Recombinant Proteins;
biosynthesis;
genetics;
pharmacology
- From:
Chinese Journal of Biotechnology
2009;25(7):1035-1041
- CountryChina
- Language:Chinese
-
Abstract:
We constructed the recombinant adenovirus vector expressing IL-24 and E1A (Ad-IL-24-E1A) and investigated the inhibition of Ad-IL-24-E1A on SMMC-7721 hepatocellular carcinoma in vitro. We amplified IL-24 gene by PCR using pAdTrack-IL-24 as template. The IL-24 gene was cloned into pAdTrack-IRES at the Bgl II and Sal I site to form pAdTrack-IL-24-IRES. E1A digested from pAdTrack-E1A was cloned into the pAdTrack-IL-24-IRES at the Xho I and EcoR V site to form the pAdTrack-IL-24-IRES-E1A. We co-transformed both pAdTrack-IL-24-IRES-E1A and pAdeasy-1 digested by Pme I and packaged to obtain Ad-IL-24-E1A. Ad-IL-24-E1A at 50 MOI infected SMMC-7721 cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay determined cell proliferation. Flow cytometry detected Cell apoptosis. The apoptotic rate of SMMC-7721 cells was 52% 48 h after infection with Ad-IL-24-E1A. The result showed that the growth of SMMC-7721 cells was significantly inhibited by Ad-IL-24-E1A at the MOI of 50.
