| Objective:Herpes simplex virus thymidine kinase gene is one of the most widely used gene therapy methods for cancer.This study was to investgate the treatment efficiency of replication defective adenovirus carrying HSV/TK gene under control of the human telomerase reverse transcriptase (hTERT) promoter and ganciclovir (GCV) on the formation of carcinomatous ascites of hepatocellular carcinoma in mice.Methods:1. Ad-hTERTp-HSV-TK was amplified,purified by using HEK 293 cells and viral titer was determined.2. The SX1 inbred strain mice were injected with H22 cell line of liver cancer and were divided into 4 groups at random. The mice in each group were given corresponding treatment after 48 hours. The therapeutic effect was evaluated by comparing the change of weights,abdominal circumference,ascites volumes,survival time and influence of the main internal organs.3. The apoptosis rates of tumor cells were detected by FCM.4. Morphological changes of tumor cells were observed by electromicroscope after treatment.Results:1. The viral titer was 1.5×1010pfu/ml after amplification and purification. 2. Ad-hTERTp-HSV-TK/GCV system can obviously inhibit the production of ascites.Theascites volume was obviously small as compared to any control group (P<0.01),prolong the survival period(P<0.01). No obvious side effect was found during the treatment.3. Apoptosis rate in Ad-hTERTp-HSV-TK/GCV group (27.12±2.12)% was significantly higher than that in other groups.4. Typical morphological changes of apoptotic cells in Ad-hTERTp-HSV-TK/GCV group were observed by electron microscope.Conclusion:Ad-hTERTp-HSV-TK/GCV system can inhibit production of ascites and prolong the survival period of mice by inducing apoptosis of hepatoma cells, which is a safe and feasible treatment for hepatoma therapy. It is promising to be used in clinical gene therapy. |
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