Construction of adenovirus carrying dual-target shRNA for Oct-4 and Survivin and its inhibitory effect on human hepatocellular carcinoma cells.
- Author:
Duanming WANG
1
;
Jinghan WANG
;
Linfang LI
;
Jingbo CHEN
;
Changqing SU
Author Information
1. Key Laboratory of Xinjiang Endemic and Ethnic Diseases, School of Medicine, Shihezi University, Shihezi 832000, Xinjiang, China.
- Publication Type:Journal Article
- MeSH:
Adenoviridae;
genetics;
metabolism;
Animals;
Apoptosis;
genetics;
Carcinoma, Hepatocellular;
pathology;
therapy;
Cell Line, Tumor;
Genetic Therapy;
Genetic Vectors;
genetics;
HEK293 Cells;
Humans;
Inhibitor of Apoptosis Proteins;
biosynthesis;
genetics;
Liver Neoplasms;
pathology;
therapy;
Mice;
Mice, Nude;
Octamer Transcription Factor-3;
biosynthesis;
genetics;
RNA, Small Interfering;
genetics;
Recombinant Proteins;
biosynthesis;
genetics;
Transfection
- From:
Chinese Journal of Biotechnology
2012;28(5):623-631
- CountryChina
- Language:Chinese
-
Abstract:
The transcriptional factor Oct-4 and Survivin are the key regulatory factors in cancer cell proliferation and mitosis. A dual cancer-specific shRNA adenovirus vector, Ad5-Dual-shRNA, targeting Oct-4 and Survivin genes was constructed by molecular cloning and recombination. After cells were infected with virus, hepatocellular carcinoma cell line EHBH-H1 was used for detecting the expression of Oct-4 and Survivin proteins by Western blotting. The viral cytotoxic effect on cancer cells was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay in vitro, and the inhibition effect on tumor xenografts was observed in nude mice. The results showed that the expression of Oct-4 and Survivin in cancer cell line EHBH-H1 could be silenced markedly by Ad5-Dual-shRNA. In MTT and animal experiments, Ad5-Dual-shRNA also represented much stronger anti-tumor effect on tumor growth than Ad5-Surv-shRNA and Ad5-Oct4-shRNA. From this research we can draw a conclusion that the cancer-specific adenovirus vector expressing dual-shRNA targeting Oct-4 and Survivin genes may provide us a more effective, specific and convenient gene therapy method.
