The construction of transferrin receptor- mediated HSV-TK gene transfer system and its effect on human hepatocellular carcinoma cells in vitro.
- Author:
Dao-feng YANG
1
;
Hui-fen ZHU
;
Guan-xin SHEN
;
De-ying TIAN
Author Information
- Publication Type:Journal Article
- MeSH: Antibodies, Monoclonal; therapeutic use; Carcinoma, Hepatocellular; therapy; Cell Line, Tumor; Ganciclovir; therapeutic use; Genetic Therapy; Humans; Liver Neoplasms; therapy; Receptors, Transferrin; immunology; Simplexvirus; enzymology; Thymidine Kinase; genetics; alpha-Fetoproteins; genetics
- From: Chinese Journal of Hepatology 2004;12(2):88-91
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the localization system involving anti-TfR monoclonal antibody (McAb) and AFP promoters and assess its effect on human hepatoma cell lines.
METHODSThe conjugate of anti-TfR McAb and polylysine (PLL) was made by SPDP and purified by molecular screen chromatography. DNA blocking test determined that the ratio of one pEBAF/tk to six Ab-PLL was the most suitable to couple them. The pEBAF/tk recombinant plasmid bearing HSV-TK gene was coupled to Ab-PLL by noncovalent bond. The pEBAF/tk was transferred into human hepatoma cell line HepG2, SMMC7721 and pulmonary cancer cell line A549 by receptor-mediated gene delivery (Ab-PLL-DNA) and liposome procedure. The growth inhibitory rates of HepG2, SMMC7721 and A549 cells were measured by MTT assay.
RESULTSThe inhibitory rates of HepG2/tk in 100 mg/L and 1 mg/L of GCV were 60.5% and 24.3%, respectively. The inhibitory rate of GCV to SMMC7721 was 23.2% in 3 days. The pulmonary cancer cell A549, A549/tk (Ab) and A549 /tk (lipo) could not be inhibited by the addition of GCV.
CONCLUSIONThe localization system employed in this paper has high specificity, effectiveness and safety for gene therapy. It would be a promising strategy for gene therapy.