The Cytotoxic Effect Of The Suicide Gene Fcy: Fur::Fur Mediated By Retrovirus Vector On The Transformed NIH3T3 Cells Induced By KSHV Tumorigenesis Gene K12

To study the cytotoxic effect of cytosine deaminase (CD)/5-fluorocytosine (5-FC) on the transformed NIH3T3 cells induced with Kaposi's sarcoma-associated herpesvirus (KSHV) tumorigenesis gene K12. Two recombinant retrovirus expressing plasmids LZRS-K12 and pLXSN-FF containing KSHV K12 gene and CD suicide gene Fcy::Fur were generated, respectively, with gene recombinant techniques. Then two cell lines named N/K12 and N/K12/FF expressing kaposin protein were obtained by stably transfecting NIH3T3 cells with the plasmid LZRS-K12 alone or together with the recombinant pLXSN-FF. By virtue of common inverse microscope, trans-electric microscope, soft-agar colony forming assay and nude mice tumorigenesis tests, the transforming ability in vitro and the tumorigenesis ability in vivo of K12 gene were identified, respectively. The growth characteristic and shape of the NIH3T3 cells transfected with K12 gene changed obviously, which showed heteronucleus and growing for multilayer for losing contact inhibition, and forming cell colony on soft-agar for lower growing dependence. In vivo experiments demonstrated that tumors could be observed on the nude mice after inoculated with NIH3T3 cells transfected with K12 gene for 2-3 weeks. Slice of the tumor tissue dyed with haematoxylin-eosin indicated...