Abstract:Objective;To investigate the repressing effects of cytosine deaminase(CD) and herpes simplex virusthymidine kinase(HSVtk) double suicide genes coexpression system on human cholangiocarcinoma QBC939 cells and QBC939 cells transplantated tumor in nude mice. Methods;CD and HSVtk double suicide genes were transfered into QBC939 cells using liposomes. After G418 selection, the positive clones of QBC939/CD+tk cells were picked up and cultured. The expression of CD and HSVtk genes was confirmedby RTPCR. In vitro, the QBC939/CD+tk cells were treated with 5Fc and/or GCV, and the cytoxicity efficacy was evaluated by microculture tetrajolium test (MTT) method. The QBC939/CD+tk cells were inoculated subcutaneously into nude mice, and when the tumors were palpable, 5Fc and GCV were injectedintraperitoneally, and the volumes of transplantated tumors were measured before and after medication. Results;Double suicide genes were stably expressed in QBC939/CD+tk cells. The repressing capability of combination of 5Fc and GCV on QBC939/CD+tk cells was more effective than that of using either 5Fc or GCV alone. The increase of cellrepressing was assaciated with increase the concentration of the prodrug. The repressing effect of combination of the 2 prodrugs on early period of transplantation tumor was obvious, even complete abolition of tumor was noted, and moreover a marked local bystander effect was observed. Conclusions;In vitro and in vivo, the cellrepressing efficacy of double suicide gene system on cholongiocarcinoma cells and the tramsplanted tumor of nude mice was significant, and the bystander effect was obvious.