为进一步探究外源性绵羊肺腺瘤病毒(exJSRV)囊膜蛋白(Env)的致病机制,本研究检测了自然感染绵羊肺腺瘤病(OPA)的绵羊肺组织中p-Akt和p-mTOR的表达定位、Akt和mTOR在肺组织中的磷酸化水平以及Beclin1在病肺组织中的表达水平,同时通过表达exJSRV-env的重组表达质粒诱导转化NIH3T3细胞,检测诱导转化的细胞中Akt和mTOR的磷酸化水平、Beclin1的表达水平.此外,在诱导转化的细胞中添加mTOR抑制剂Rapamycin后,检测其对Beclin1表达的影响.结果显示,p-Akt和p-mTOR主要表达于自然感染OPA病变肺组织的肺泡上皮细胞和肿瘤细胞中,与健康肺组织比较,在自然感染OPA病变肺组织中的二者磷酸化水平升高显著(p＜0.05),而Beclin1在自然感染OPA病肺组织中的表达降低显著.p-mTOR在转染env的NIH3T3细胞中的磷酸化水平极显著高于对照组细胞(p＜0.01),Beclin1在转染env的NIH3T3细胞中的表达量极显著低于正常的NIH3T3细胞(p＜0.01).而添加mTOR抑制剂Rapamycin后转染env的NIH3T3细胞与对照组细胞相比,Beclin1的表达水平显著上调(p＜0.05).结果表明,自然感染OPA的病肺组织和转染env的NIH3T3细胞的Akt/mTOR信号通路被激活,并通过该通路抑制自噬,提示exJSRV-env可能通过抑制病变细胞的自噬促进肿瘤的形成.本研究为进一步探索JSRV Env诱导转化细胞及与细胞自噬之间的关系的研究奠定了基础.%In order to explore the pathogenesis of exogenous Jaagsiekte sheep retrovirus envelope protein (exJSRV-env),we detected the locations of phospho-Akt (p-Akt) and phospho-mTOR (p-mTOR) in Ovine pulmonary adenocarcinoma (OPA) in sheep lung and the relative expressions of p-Akt,p-mTOR and Beclin1 in sheep lung and in NIH3T3 cells after transfecting with pcDNA4/myc-His/exJSRV-env which expressed exJSRV-env,and we also detected the level of Beclin1 after treated with rapamycin.The results showed that infected lung cells expressed p-Akt and p-mTOR in alveolar epithelial cells and tumor cells,the activation of p-Akt and p-mTOR in OPA sheep lung were higher than normal sheep lung and the expression of Beclin1 in OPA sheep lung was lower than normal lung.The relative expression of p-mTOR in NIH3T3 cells transfected with pcDNA4/myc-His/ exJSRV-env was higher than the control cells,and the level of Beclin1 in the transfected NIH3T3 cells was lower than the control cells.The level of Beclin1 was increased when NIH3T3 cells were treated with rapamycin.We conclude that Akt/mTOR signal transduction pathway was activated in OPA lung cells and in the NIH3T3 cell stransfected with.JSRV Env decreased Beclin1 by mTOR pathway.We provide a platform for the pathogenesis of JSRV Env and the relationship between JSRV Env and autophagy.
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