| The malignant tumor is the disease of endangering the human health badly. . In vivo, transfer of HSV1-tk gene plus GCV combined with Liuwei Dihuang Decoction have synergistic antitumor effects.So the theory and pratical bases are provided in order to how to improve the effects of combining therapy by explaining this mechanism .Objective: To construct a new cell stain by transfect tk gene to H22 cell stain and name it H22/tk. Then use it for treating tumor in vivo and observe the curativeeffect of treatment by combining Liuwei Dihuang Bolus and GCV. And study the mechanism by immunohistochemical and elisa method. Provide requirement for gene therapy of tumor by combining traditional Chinese and western medicine.Methods: Based on PT67/tk cell stain has been construction by DNA recombinant technique, then determining vival titer by infecting N1H 3T3 cells, H22 was infected with the recombinant retrovirus. The positive clones were obtained after G418 selection and named H22/tk. The morphological changes after GCV treatment were observed with microscope. The liver tumor model on the KM mice were made in vivo by inoculate the cell mix whick comprise 80% H22 and 20% H22/tk. Give a treatment by injecting pro-drug GCV, and combined with Liuwei Dihuang Bolus. Observe the weight of tumor after 14 days and histopathological expression of the tumors were observed in every group. Observe the expression of Cx43 by immunohistochemical technic. Observe the concentration of IFN and TNF in serum of tumor-endured mice .Results: The titer of the virus which is excreted by PT67/tk was 2× 10~5cfu/mL. In vitro, through morphological observation. H22/tk cells were highly sensitive to GCV. In vivo, mice with tumor received combined therapy developed tumors more slowly, when compared with mice treated with HSV-tk/GCV alone, the average tumor weights were much less in the combination group, which was significant by comparing with that in... |
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