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


CONCLUSIONS
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.
Wang
Chun-ting
CT
State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
Zhang
Peng
P
Peng
Feng
F
Ruan
Xu-zhi
XZ
Yang
Han-shuo
HS
D000818:Animals; D015153:Blotting, Western; D049109:Cell Proliferation; D051379:Mice; D041681:NIH 3T3 Cells; D009363:Neoplasm Proteins / Q000235:genetics / Q000502:physiology*; D010957:Plasmids; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D009363:Neoplasm Proteins