The Studies Of Tumorigenigcity Of JSRV-Env In Rats

OBJECTIVE:To explore the tumorigenic role of JSRV-Env, and the affact for the animal’s body of physiological parameters and related gene.METHODS:The eukaryotic expression plasmid pcDNA3.1-env is transfected into young rats with vivo transfection reagent through tail vein injection method. After21days, using RT-PCR and immunohistochemical staining method to detect the eukaryotic expression plasmid pcDNA3.1-env expression in rats. After that, we determinated the clinical indicators of the rats which were been transtected within six months, and detected the dynamic changes of flow cytometry in peripheral blood lymphocyte subsets and lymphocyte apoptosis every30days. At the same time, we should conduct the blood gas analysis and detection of tumor markers, and detected the mutations of the organization of rat lung K-ras gene, P53gene used the SSCP method to detect. Also observed the pathological changes of the organizations through the production of pathological.RESULTS:After transfected21days, eukaryotic expression plasmid pcDNA3.1-env was expressde in lung tissue of rats. the rats which have been transfected for180days, the level of CD4+and CDS+T lymphocyte show that they didn’t have significant changes in the whole process.The rusult of Lymphocyte apoptosis show that the level of apoptotic cells did not occur significant apoptosis。pCO2, pO2, Hct of BE (ecf) and other sixteen indicators compared with the control group, there is no significant changes。CEA and AFP were normal, the transfected group compared with the control group, they did not have significant changes。K-ras gene and P53gene didn’t occur mutation. Histopathology did not find the characteristic pathological changes.CONCLUSIONS:In this study, rats were detected JSRV-env gene mRNA expression, at the same time, env gene was expressed, which prove that JSRV-env gene has been successfully transfected into the cytoplasm of lung cells, the Env glycoprotein of JSRV not only play the role of viruses invasive into cells, but also got have the similar role of viral oncoprotein, it can directly stimulate cell transformation, which laid the foundation for the next tumorigenic effect。In this study, rats were transfected by JSRV pcDNA3.1-env eukaryotic expression plasmid, through the detection of blood-gas analysis, detection of lymphocyte subsets, detection of lymphocyte apoptosis, tumor markers, gene mutations detection and histopathology tests, which compare with the control group, ther were not found significant changes, the results confirm that the JSRV-env didn’t produce tumorigenic effect for rats. JSRV-env didn’t integrated into the rat genome may be the main reason, thus not play a tumorigenic role. Probably because of the animal species difference, or the time is not enough.