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采用二维差异凝胶电泳-质谱联用技术鉴定肺鳞状细胞癌中肿瘤相关蛋白靶点的蛋白质组学方法。

Proteomics approaches for identification of tumor relevant protein targets in pulmonary squamous cell carcinoma by 2D-DIGE-MS.

作者信息

Lihong Hao, Linlin Gong, Yiping Guo, Yang Song, Xiaoyu Qi, Zhuzhu Guan, Xiaohan Yang, Xin Zhou, Liyan Xue, Shujuan Shao

机构信息

Key Laboratory of Proteomics, Dalian Medical University, Dalian, Liaoning Province, P.R. China.

Department of Pathology, Cancer Institute and Hospital, Chinese Academy of Medical Sciences (CAMS) and Peking Union Medical College (PUMC), Beijing, P.R. China.

出版信息

PLoS One. 2014 Apr 16;9(4):e95121. doi: 10.1371/journal.pone.0095121. eCollection 2014.

Abstract

Potential markers for progression of pulmonary squamous cell carcinoma (SCC) were identified by examining samples of lung SCC and adjacent normal tissues using a combination of fluorescence two-dimensional difference gel electrophoresis (2D-DIGE), matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and electrospray ionization quadrupole-time of flight mass spectrometry (ESI-Q-TOF). The PANTHER System was used for gel image based quantification and statistical analysis. An analysis of proteomic data revealed that 323 protein spots showed significantly different levels of expression (P ≤ 0.05) in lung SCC tissue compared to expression in normal lung tissue. A further analysis of these protein spots by MALDI-TOF-MS identified 81 different proteins. A systems biology approach was used to map these proteins to major pathways involved in numerous cellular processes, including localization, transport, cellular component organization, apoptosis, and reproduction. Additionally, the expression of several proteins in lung SCC and normal tissues was examined using immunohistochemistry and western blot. The functions of individual proteins are being further investigated and validated, and the results might provide new insights into the mechanism of lung SCC progression, potentially leading to the design of novel diagnostic and therapeutic strategies.

摘要

通过结合荧光二维差异凝胶电泳(2D-DIGE)、基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)和电喷雾电离四极杆飞行时间质谱(ESI-Q-TOF),对肺鳞状细胞癌(SCC)样本及相邻正常组织进行检测,从而确定肺鳞状细胞癌进展的潜在标志物。使用PANTHER系统进行基于凝胶图像的定量分析和统计分析。蛋白质组学数据分析显示,与正常肺组织中的表达相比,323个蛋白质点在肺SCC组织中的表达水平存在显著差异(P≤0.05)。通过MALDI-TOF-MS对这些蛋白质点进行进一步分析,鉴定出81种不同的蛋白质。采用系统生物学方法将这些蛋白质映射到参与众多细胞过程的主要途径,包括定位、运输、细胞成分组织、凋亡和繁殖。此外,使用免疫组织化学和蛋白质印迹法检测了肺SCC和正常组织中几种蛋白质的表达。正在进一步研究和验证单个蛋白质的功能,其结果可能为肺SCC进展机制提供新的见解,有可能促成新型诊断和治疗策略的设计。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03d7/3989308/f6ee2c054f85/pone.0095121.g001.jpg

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