Primary Tumor Xenografts of Human Lung Adeno and Squamous Cell Carcinoma Express Distinct Proteomic Signatures
Primary Tumor Xenografts of Human Lung Adeno and Squamous Cell Carcinoma Express Distinct Proteomic Signatures
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DOI：
10.1021/pr100491e
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发表时间：
2011-01-01
影响因子：
4.4
通讯作者：
Moran, Michael F.
中科院分区：
文献类型：
--
作者：
Wei, Yuhong;Tong, Jiefei;Moran, Michael F.
非小细胞肺癌（NSCLC）占肺癌的80%。非小细胞肺癌最常见的亚型是腺癌（ADC）和鳞状细胞癌（SCC），两者加起来约占90%。将10例切除的NSCLC患者肿瘤（5例ADC和5例SCC）直接导入严重免疫缺陷（NOD-SCID）小鼠，采用标准组织学和免疫组织化学（INC）以及蛋白质组学分析所产生的异种移植肿瘤。质谱（MS）方法包括一维和二维LC-MS/MS，以及多重选择反应监测（SRM，或MRM），用于鉴定和量化异种移植物蛋白质组。蛋白质谱的分层聚类区分了ADC和SCC亚型。178种蛋白的差异表达，包括一组全面的中间丝角蛋白，被发现构成了与NSCLC亚型相关的独特蛋白质组学特征。表皮生长因子受体（EGFR）在ADC和SCC异种移植物中表达，并通过磷酸化酪氨酸谱分析、Western blot分析和SRM测量EGFR水平和突变分析来评估EGFR网络激活情况。多重SRM/MRM方法在单次LC-MS/MS运行中提供了几种角蛋白、EGFR和plakophilin-1的相对定量。SRM和MS/MS光谱计数的蛋白质定量具有较好的动态范围和重复性，但与免疫组化和免疫印迹等正交方法一致。这些发现说明了在肿瘤病理学中开发一个基于ms的综合平台的潜力，以更好地分类和潜在地治疗非小细胞肺癌患者。
Nonsmall cell lung carcinoma (NSCLC) accounts for 80% of lung cancers. The most prevalent subtypes of NSCLC are adenocarcinoma (ADC) and squamous cell carcinoma (SCC), which combined account for approximately 90%. Ten resected NSCLC patient tumors (5 ADC and 5 SCC) were directly introduced into severely immune deficient (NOD-SCID) mice, and the resulting xenograft tumors were analyzed by standard histology and immunohistochemistry (INC) and by proteomics profiling. Mass spectrometry (MS) methods involving 1- and 2-dimensional LC-MS/MS, and multiplexed selective reaction monitoring (SRM, or MRM), were applied to identify and quantify the xenograft proteomes. Hierarchical clustering of protein profiles distinguished between the ADC and SCC subtypes. The differential expression of 178 proteins, including a comprehensive panel of intermediate filament keratin proteins, was found to constitute a distinctive proteomic signature associated with the NSCLC subtypes. Epidermal growth factor receptor (EGFR) was expressed in ADC and SCC xenografts, and EGFR network activation was assessed by phosphotyrosine profiling by Western blot analysis and SRM measurement of EGFR levels, and mutation analysis. A multiplexed SRM/MRM method provided relative quantification of several keratin proteins, EGFR and plakophilin-1 in single LC-MS/MS runs. The protein quantifications by SRM and MS/MS spectral counting were associated with superior dynamic range and reproducibility but were otherwise consistent with orthogonal methods including IHC and Western immuno blotting. These findings illustrate the potential to develop a comprehensive MS-based platform in oncologic pathology for better classification and potentially treatment of NSCLC patients.