Title: Therapeutic effect of triple regulated adenovirus vector expressing mda7/IL24 on liver cancer in vitro
Abstract: Objective: To construct an E1A-deleted 24-bp triple regulated replicative adenovirus vector SG600/interleukin24(IL24),which was driven by both hTERT promoter and HRE promoter.The level of IL24 in liver cancer cells was determined and the replication capacity of SG600/IL24 and its killing effects on liver cancer cells were observed.Methods: SG600-IL24 vector was constructed using DNA cloning and recombination techniques.The IL24 gene expression in liver cancer cell lines SMMC-7721 and BEL-7404 and normal cell line BJ was detected by ELISA assay.The replications of SG600/IL-24 in different cell lines were determined by evaluating TCID50(50% tissue culture infectious dose) at 48 and 96 h.In vitro cell-killing effects of SG600/IL24 on the three liver cancer cell lines were analyzed by MTT assay and CPE(cytopathic effect) staining method at different MOI values.Results: IL24 was over-expressed in both SMMC-7721 and BEL-7404 cells but was weakly expressed in BJ cells.At 48 and 96 h post infection the replication of SG600/IL-24 were 794 and 7940 folds in SMMC-7721 cells;622 and 7 810 folds in BEL-7404 cells;20 and 200 folds in BJ cells.MTT assay showed that the MOI values of SG600/IL24 for killing 50% and 90% cells were 0.3 and 5 for SMMC-7721 cells;3 and 20 for BEL-7404 cells;50 and 150 for BJ cells.CPE staining demonstrated that SG600/IL24 had significant killing effects on both liver cancer cells SMMC-7721 and BEL-7404 but had no significant influence on BJ cells.The cell-killing capability of SG600/IL24 was superior than that of replicative adenovirus ZD55/IL24 and non replicative adenovirus Ad-IL24.Conclusion: After SMMC-7721 and BEL-7404 liver cancer cells are infected with SG600/IL24 at high efficiency,the virus replication is active and the expression of IL24 increases greatly.SG600/IL24 has specific cell-killing effects on the two liver cancer cell lines but has no significant influence on normal cells.This study provides a basis for further investigating the effect of SG600/IL24 on liver cancer in vivo.
Publication Year: 2008
Publication Date: 2008-01-01
Language: en
Type: article
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