State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, Sichuan Province, China.
J Cell Biochem. 2009 Dec 1;108(5):1211-9. doi: 10.1002/jcb.22353.
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.
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。