本试验旨在进一步全面研究绵羊肺腺瘤病毒(JSRV)囊膜蛋白ENV的功能。使用本试验室已构建好的真核表达质粒pcDNA3.1(+)/exJSRV-env转染NIH3T3细胞,并将此NIH3T3细胞接种到裸鼠体内;重组质粒经尾静脉注射到幼龄BALB/c小鼠体内。结果显示,真核表达质粒pcDNA3.1(+)/exJSRV-env引起NIH3T3细胞发生转化,转染细胞接触抑制性消失,并形成大量的转化灶;pcDNA3.1(+)/exJSRV-env转染的NIH3T3细胞接种裸鼠使裸鼠致瘤,即引起NIH3T3细胞发生癌变。JSRVenv基因在小鼠体内表达,肝脏细胞出现大面积弥散性坏死,脾脏白髓严重坏死,心肌细胞明显颗粒样变性,肺间质细胞增生,肾小管上皮细胞轻度颗粒样变性。结果表明,绵羊肺腺瘤病毒囊膜蛋白可以引起鼠NIH3T3细胞发生转化和癌变,使小鼠各脏器出现不同程度的病变。本试验为进一步探索JSRV ENV的致瘤机制奠定基础。
To further explore the function of the Inner Mongolia strains of JSRV envelope protein, using the constructed eukaryotic expression plasmid pcDNA3. 1 (+)/exJSRV-env to transfect NIH3T3 cells in vitro ;the recombinant plasmid was injected into the tail vein of young BALB / c mice in vivo. The eukaryotic expression plasmid pcDNA3.1 (+)/exJSRV-env caused the transfor- mation of NIH3T3 cells, the contact inhibition of the transfected cells were disappeared and formed a large number of loci;the pcDNA3.1 (+)/exJSRV-env plasmid transfected NIH3T3 cells were inoculated into nude mice for tumorigenicity,JSRV-env caused the NIH3T3 cells to undergo carcinogenesis. The JSRV env gene was expressed in mice,the large area of the liver cells displayed the diffuse necrosis, splenic white pulp with serious necrosis, the myocardial cells with significantly granular degeneration and the kidney cells with mild granular degeneration, pulmonary interstitial cell hyperplasia. This experiment showed that the JSRV envelope protein could induced the trans- formation and carcinogenesis of the NIH3T3 cells and displayed varying degrees of pathological changes in mice organs, this experiment lays the foundation for further study on the tumorigenic mechanism of JSRV ENV.