Search

Background & Objective: Adenovirus vector-mediated herpes simplex virus thymidine kinase gene (ADV-TK) transfer is one of the major gene therapy strategies for tumor. This study was designed to determine the in vitro anti-tumor efficiency of ADV-TK, a recombinant construct previously developed in our laboratory., Methods: Herpes simplex virus thymidine kinase (TK) gene was transduced into 14 types of cultured tumor cells with different histological origins using adenovirus vector followed by ganciclovir (GCV) medication. The killing efficiency was measured using MTT assay., Results: At the dosage of 1x10(9) viral particles/per well in the presence of 100 microg/ml GCV, ADV-TK/GCV caused effective killing of 11 out of total 14 types of tumor cells with a rate higher than 74%, the other 3 tumor cells, laryngeal epithelial cancer cells (Hep-2), hepatic cancer cells (Bel7402), and human colon cancer cells (HCT-8) were less sensitive to the ADV-TK/GCV treatment with the killing rates of 55.3%+/-2.0%, 61.3%+/-2.0%, and 63.7%+/-2.5%, respectively. Except for Hep-2, the killing efficiency caused by ADV-TK/GCV treatment was similar to that caused by cisplatin at a dosage equal to the in vivo peak concentration (5 microg/ml) in tissue., Conclusion: ADV-TK is highly efficient for active killing of tumor cells in vitro and is promising for future clinical application.

Objective: To detect the expression of recombinant human bone morphogenetic protein7 (rhBMP7) in skeletal muscle satellite cells (SMSCs) with rhBMP7 gene transfection mediated by retroviral vector., Methods: rhBMP7 gene was reconstructed in retroviral vector and transferred into packaging cells PT67 via liposome reagent, with the positive cell clones selected with G418. In vitro cultured SMSCs were transfected with the virus granules secreted by PT67 cells and followed by G418 selection. Reverse transcriptase-polymerase chain reaction was utilized for analysis for rhBMP7 mRNA in the transfected cells., Results: rhBMP7 retrovirus vector was successfully constructed and transferred into PT67 cells, and abundant mRNA expression of rhBMP7 was observed in the skeletal muscle satellite cells transfected with the virus and selected with G418., Conclusion: rhBMP7 gene can be transferred into the skeletal muscle satellite cells via retroviral vector to yield effective rhBMP7 mRNA expression.

Objective: To enhance the therapeutic effects on nasopharyngeal carcinoma by combining treatment with selectively replicating adenovirus and IL12., Methods: Replicating adenovirus with mouse IL12 gene insert (CNHK200-mIL12) was constructed to transfect nasopharyngeal carcinoma cell lines CNE3 and 915. Adenovirus hexon was detected by immunohistochemical staining and flow cytometry (FCM), and mIL12 expression examined by enzyme-linked immunosorbent assay (ELISA). The replication rates of CNHK200-mIL12 and dl1520 was determined by 50% tissue culture infectious dose (TCID50)., Results: Twenty-four hours after transfection with CNHK200-mIL12, most of cells were positive for adenovirus hexon and FCM demonstrated increased positivity rates of 39% and 4% among CNE3 and 915 cells respectively. It was observed that CNHK200-mIL12 replication increased by 1 000 folds with mIL12 expression level reaching as high as 84.5+/-4.6 ng in CNE3 cells and 75.6+/-3.4 ng in 915 cells as determined 72 h after transfection with 1x10(5) PFU CNHK200-mIL12 into 1x10(4) cells., Conclusion: CNHK200-mIL12 can replicate in vitro in nasopharyngeal carcinoma cells (dl1520 cells, for instance) with high mIL12 expression, which suggests that CNHK200-mIL12 may potentially be used to treat nasopharyngeal carcinoma.