PI3K/Akt信号转导通路在ALV-J感染中作用的初步研究

doi:10.3864/j.issn.0578-1752.2011.16.018

中国农业科学 ›› 2011, Vol. 44 ›› Issue (16): 3446-3453.doi: 10.3864/j.issn.0578-1752.2011.16.018

PI3K/Akt信号转导通路在ALV-J感染中作用的初步研究

冯少珍, 李娇, 吴晓婵, 曹伟胜, 廖明

华南农业大学兽医学院/农业部动物疫病防控重点开放实验室

收稿日期:2011-04-14 修回日期:2011-05-11 出版日期:2011-08-15 发布日期:2011-06-22
通讯作者: 通信作者廖明，Tel：020-85280240；E-mail：mliao@scau.edu.cn。曹伟胜，E-mail：caoweish@scau.edu.cn
作者简介:冯少珍，E-mail：fengshaozhen111@hotmail.com
基金资助:
NSFC-广东联合基金（U0831002）、国家自然科学基金（30771612）、广东省自然科学基金（10451064201005432）、国家博士后科学基金（20100470929）和广东省科技计划项目（2009A020101006）

The Function of PI3K/Akt Signal Pathway During ALV-J Infection in DF-1 Cells

FENG Shao-Zhen, LI Jiao, WU Xiao-Chan, CAO Wei-Sheng, LIAO Ming

华南农业大学兽医学院/农业部动物疫病防控重点开放实验室

Received:2011-04-14 Revised:2011-05-11 Online:2011-08-15 Published:2011-06-22
Contact: Shao-zhen FENG

摘要/Abstract

摘要： 【目的】探讨ALV-J在宿主细胞中复制与PI3K/Akt信号转导通路的关系。【方法】将血管瘤病变型ALV-J毒株HN06和骨髓瘤病变型ALV-J毒株NX0101分别感染DF-1细胞，通过Western blot、Real-time PCR、IFA和ELISA等方法，观察细胞Akt蛋白磷酸化水平、病毒RNA表达水平和病毒蛋白表达水平等指标。【结果】HN06株和NX0101株在体外细胞中复制水平有差异。HN06株的早期感染可引起Akt转导通路的活化，病毒引起的Akt磷酸化具有病毒滴度依赖性，而且能被PI3K特异性抑制剂LY294002所抑制，表明HN06株诱导的Akt活化是PI3K途径依赖的。LY294002可在病毒感染早期呈剂量依赖性地显著降低受染细胞中HN06 RNA水平、囊膜蛋白水平和细胞培养物上清中的病毒粒子含量。【结论】PI3K/Akt信号转导通路活化对HN06株在细胞感染早期具有重要的作用，该结果与已报道的有关细胞PI3K/Akt信号转导通路参与NX0101株的早期感染的结论一致。本研究为进一步阐明ALV-J入侵宿主细胞和复制的精确机制等研究奠定了基础。

关键词: J亚群禽白血病病毒, 血管瘤, 髓细胞瘤, 感染早期, PI3K/Akt

Abstract: 【Objective】 Given the biological importance of the PI3K/Akt pathway in virus infection, this experiment investigated whether or not the subgroup J avian leukosis virus (ALV-J) strain HN06 infection in DF-1 cells is correlated with the activity of Akt. 【Method】After inoculation of ALV-J strain HN06 which mainly induces hemangiomas and strain NX0101 which induces myelocytomas in DF-1 cells, respectively, the expression levels of Akt phosphration, viral RNA transcription and viral protein synthesis were analyzed.【Result】 Results showed that the replication rates in vivo of ALV-J strains HN06 and NX0101 were different. HN06 infection led to increased Akt phosphorylation at a very early stage which was virus titer-dependent. LY294002, a PI3K-specific inhibitor, could suppress HN06 induced Akt phosphorylation, indicating that Akt phosphorylation was PI3K-dependent. Moreover, in the presence of LY294002, viral RNA transcription level, viral envelope protein ENV expression and virons secreted in supernatants were suppressed significantly. The PI3K/Akt activation profile and function of HN06 were similar to that of NX0101 reported. 【Conclusion】These data suggest that PI3K/Akt signaling pathway plays an important role in ex vivo ALV-J replication, although the precise mechanism remains under investigation.

Key words: Subgroup J avian leukosis virus, Hemangioma, myelocytomas, Virus early infection, PI3K/Akt signal pathway

中图分类号:

冯少珍, 李娇, 吴晓婵, 曹伟胜, 廖明. PI3K/Akt信号转导通路在ALV-J感染中作用的初步研究[J]. 中国农业科学, 2011, 44(16): 3446-3453.

FENG Shao-Zhen, LI Jiao, WU Xiao-Chan, CAO Wei-Sheng, LIAO Ming. The Function of PI3K/Akt Signal Pathway During ALV-J Infection in DF-1 Cells[J]. Scientia Agricultura Sinica, 2011, 44(16): 3446-3453.

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PI3K/Akt信号转导通路在ALV-J感染中作用的初步研究

The Function of PI3K/Akt Signal Pathway During ALV-J Infection in DF-1 Cells

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