Studies On The Influence Of JSRV-Env Recombinant Plasmids On The Expression Of Several Cell Growth Factors Of Three Kinds Of Murine Cells

Sheep pulmonary adenomatosis (Ovine pulmonary adenomatosis, OPA or Sheep pulmonary adenomatosis, SPA) pathogenic factor-sheep pulmonary adenoma virus (JSRV), is not yet entirely researched clearly of OPA complex pathogenesis, and regarding to tumor-associated cytokines in the pathogenesis association of OPA research data rarely reported. The study found that four cytokine VEGF, EGFR, TGF-a and EGF with tumor angiogenesis and cell proliferation and more than links were closely related. This study was to detect expression of four cytokine in target cells which were transfected with JSRV-Env recombinant plasmid (pEGFP-C1-env), and revealed the correlation both them and OPA tumor, and provided OPA address the above cytokine gene targeted therapy program with certain theoretical basis.This study selected method of liposome to make the plasmid (pEGFP-C1-env) transiently transfected TC-1, TC-1-Hyal2 and NIH 3T3 cells. Then bying the method of RT-PCR and Western blot to detect Env in three cell expression, and while using confocal microscopy determined the localization of Env in the three cell lines. Finally, after three cells were transfected, applying methods of PCR and ELISA to detect VEGF, EGFR, TGF-a and EGF of mRNA and protein levels expression. Conclusions were as follows:(1) Using liposome successfully maked plasmid pEGFP-C1-env transiently transfected with the three cell, and each of gene and protein expression levels of Env were verified.(2) VEGF, EGFR and TGF-a that expression levels of mRNA and protein both elevated, the results suggested that JSRV-Env could raise above cytokines.(3) VEGF, EGFR in NIH3T3 simultaneously displayed overexpression, the result prompted that both could be in JSRV-Env transfection vitro cell or OPA tumors existed positive correlation and during tumorigenesis played a joint catalytic role.(4) Respectively detecting two factors of EGF and EGFR expression in TC-1, TC-1-Hyal2 cells, the EGFR test results showed that the factor in two cells existed low expression phenomenon, the EGF test results prompted that the factor in two cells existed not expression phenomenon.In summary, JSRV-Env could upregulate cytokine VEGF, EGFR and TGF-a expression, this suggested that both them had a certain correlation with the OPA occur. This study for further clarified the correlation between tumor-associated cytokines and OPA occurred and its molecular mechanism provided important information,same time it could provide OPA and BAC diseases of genetic targeted treatment with an important theoretical basis.