[Effects of the novel tumor/testis antigen, Biot2, on proliferation of NIH3T3 cell].

AIM: To study the biological effects of the novel tumor/testis antigen Biot2 on proliferation of NIH3T3 cell and to identify the preliminary biological functions of it. METHODS: The Biot2 gene was amplified by RT-PCR and cloned into the eukaryotic expression vector pcDNA3.1(+) to construct the pcDNA3.1-Biot2 plasmid. The recombinant plasmid pcDNA3.1-Biot2 was transfected into NIH3T3 cell by lipofectamine 2000, and then the positive clones were screened by G418. The expression of pcDNA3.1(+) and Biot2 mRNA in positive clones were detected by RT-PCR, Realtime RT-PCR and Western blot, respectively. The cell growth curves were measured by MTT assay to study the changes of proliferation of the NIH3T3 cells stably transfected by pcDNA3.1-Biot2 plasmid and the controls. RESULTS: The eukaryotic expression vecter pcDNA3.1-Biot2 was successfully constructed. The expression of pcDNA3.1(+) vecter was found by RT-PCR. Biot2 mRNA transcription and protein were detected by Realtime RT-PCR and Western blot respectively. The growth rate of NIH3T3 cells stably transfected by the recombinant plasmid was faster than that of the controls. CONCLUSION: The stable pcDNA3.1-Biot2 transfected NIH3T3 cell line is successfully established, and the proliferation of the cells stadly transfected is promoted, which will provide experimental basis for further study on biotherapy using Biot2 target antigen.