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hPNAS4 诱导 A549 人肺腺癌细胞凋亡中蛋白质表达谱的改变。

Altered protein-expressing profile in hPNAS4-induced apoptosis in A549 human lung adenocarcinoma cells.

机构信息

State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, Sichuan Province, China.

出版信息

J Cell Biochem. 2009 Dec 1;108(5):1211-9. doi: 10.1002/jcb.22353.

Abstract

Human PNAS4 (hPNAS4) is a recently identified pro-apoptosis gene, which is able to induce apoptosis in A549 human lung adenocarcinoma cells following its overexpression. In this work, we investigated the changes of protein profile in hPNAS4-induced apoptosis in A549 cells through proteomic strategy consisting of two-dimensional electrophoresis (2-DE) coupled with MALDI-Q-TOF mass spectrometry. A total of 20 different proteins with more than 3.0-fold change in expression, including 5 up-regulated and 15 down-regulated proteins were successfully identified by database search. The mRNA transcription levels of the different proteins were further examined by RT-PCT. Functional analyses showed these different proteins are involved in diverse biological processes including metabolism, proteolysis, signal transduction, apoptosis, and redox regulation. Two essential apoptosis-associated protein, annexin A1 and prothymosin alpha, were confirmed by Western blot and showed consistent changes with proteomic detection. Our data provide molecular evidence and possible associated pathway in hPNAS4-induced apoptosis through proteomic strategy, which should be contributed to further investigation on biological function of hPNAS4.

摘要

人 PNAS4(hPNAS4)是最近发现的促凋亡基因,其过表达后可诱导 A549 人肺腺癌细胞凋亡。本研究通过二维电泳(2-DE)联合 MALDI-Q-TOF 质谱分析的蛋白质组学策略,研究 hPNAS4 诱导 A549 细胞凋亡过程中的蛋白质谱变化。通过数据库搜索,成功鉴定出 20 种差异表达蛋白,其中包括 5 种上调蛋白和 15 种下调蛋白,其表达水平变化超过 3.0 倍。进一步通过 RT-PCR 检测了不同蛋白的 mRNA 转录水平。功能分析表明,这些差异蛋白参与多种生物学过程,包括代谢、蛋白水解、信号转导、凋亡和氧化还原调节。Western blot 进一步验证了两种重要的凋亡相关蛋白 annexin A1 和 prothymosin alpha,其表达变化与蛋白质组学检测结果一致。本研究通过蛋白质组学策略为 hPNAS4 诱导的凋亡提供了分子证据和可能的相关途径,这有助于进一步研究 hPNAS4 的生物学功能。

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