谷氨酰胺激活PI3K/Akt信号通路促进痘苗病毒复制  

Glutamine Activates PI3K/Akt Signaling Pathway to Promote Replication of Vaccinia Virus

作　　者：许婉彬 王娇娇[1] 陈健华[2] 周秀梅[1] 王如伟 方玲 阎辉[4] 

机构地区：[1]浙江理工大学生命科学学院新元医学与生物技术研究所,杭州310008 [2]上海交通大学医学院附属仁济医院,上海市肿瘤研究所,上海200032 [3]浙江康恩贝制药股份有限公司,杭州310018 [4]浙江省医学科学院,杭州310013

出　　处：《中国细胞生物学学报》2017年第8期1026-1032,共7页Chinese Journal of Cell Biology

基　　金：康恩贝–浙理工中美院士工作站项目(批准号:116129A4J14365)资助的课题~~

摘　　要：谷氨酰胺(glutamine,Gln)在细胞生长和代谢过程中起重要作用,能够激活磷脂酰肌醇-3-激酶(phosphatidylinositol-3-kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)/哺乳动物雷帕霉素蛋白(mammalian target of rapamycin,m TOR)信号通路。多种病毒能通过PI3K/Akt信号通路维持细胞生存和抑制细胞凋亡,维持病毒的感染。为了探讨Gln在痘苗病毒(vaccinia virus strain western reserve,WR)感染人肺癌细胞A549过程中的调控作用及其潜在机制,该文通过蛋白免疫印迹(Western blot)检测PI3K/Akt信号通路蛋白质磷酸化水平,流式细胞术检测细胞阳性率,结晶紫染色检测WR滴度,实时定量PCR(quantitative Real-time PCR,q PCR)检测WR基因E3L和A46R m RNA水平。结果显示,Gln处理后PI3K/Akt信号通路蛋白质磷酸化水平显著升高。抑制PI3K/Akt信号通路后,细胞阳性率显著降低(P<0.05),WR滴度显著降低(P<0.05),WR基因E3L和A46R m RNA水平显著降低(P<0.05)。该研究结果表明,Gln通过激活PI3K/Akt信号通路从而促进A549细胞中WR的复制。Glutamine（Gln） plays an important role in cell growth and metabolism. It can activate phosphatidylinositol-3-kinase（PI3K）/protein kinase B（Akt）/mammalian target of rapamycin（m TOR） signaling pathway. A variety of viruses can maintain cell survival through the PI3K/Akt signaling pathway, and inhibit cell apoptosis and maintain the virus infection. This study mainly investigated the role of Gln in the process of vaccinia virus strain western reserve（WR） infection and its potential mechanism. The protein levels of PI3K/Akt signaling pathway were measured by Western blot. The cell positive rate was determined by fl ow cytometry. The WR titer was assessed by crystal violet dyeing test. The m RNA levels of gene E3 L and A46 R were determined by q PCR（quantitative Real-time PCR）. The results showed that Gln significantly promoted the levels of proteins in PI3K/Akt signaling pathway. Compared with normal Gln group, PI3K/Akt signaling pathway inhibitors significantly reduced the cell positive rate, the titer and the m RNA levels of E3 L and A46R（P〈0.05）. The results showed that glutamine could promote the WR replication by activating PI3K/Akt signaling pathway in A549 cell.

关 键 词：谷氨酰胺 痘苗病毒 PI3K/AKT信号通路 哺乳动物雷帕霉素靶蛋白 

分 类 号：R373[医药卫生—病原生物学]