Identification of Candidate Biomarkers for Early Detection of Human Lung Squamous Cell Cancer by Quantitative Proteomics
Identification of Candidate Biomarkers for Early Detection of Human Lung Squamous Cell Cancer by Quantitative Proteomics
复制标题
通过定量蛋白质组学鉴定用于早期检测人肺鳞状细胞癌的候选生物标志物
DOI：
10.1074/mcp.m111.013946
复制
发表时间：
2012-06-01
影响因子：
7
通讯作者：
Xiao, Zhi-Qiang
中科院分区：
文献类型：
--
作者：
Zeng, Gu-Qing;Zhang, Pang-Fei;Xiao, Zhi-Qiang
为了发现新的早期检测人肺鳞状细胞癌（LSCC）的生物标志物，探索LSCC可能的癌变机制，利用激光捕获微解剖纯化的正常支气管上皮（NBE）、鳞状化生（SM）、非典型增生（AH）、原位癌（CIS）和浸润性LSCC。结果，鉴定出102个差异表达蛋白，并通过Western blotting选择性验证了3个在致癌过程中表现进行性表达变化的差异蛋白（GSTP1、HSPB1和CKB）。采用免疫组化方法检测3种蛋白在独立的石蜡包埋档案标本（包括支气管上皮癌变的各个阶段组织）中的表达，并通过受体工作特征分析评估其早期检测LSCC的能力。结果表明，三种蛋白的结合可以很好地区分NBE与肿瘤前病变（SM、AH和CIS）和浸润性LSCC，区分NBE与肿瘤前病变的灵敏度为96%，特异性为92%，区分NBE与浸润性LSCC的灵敏度为100%，特异性为98%，区分肿瘤前病变与浸润性LSCC的灵敏度为92%，特异性为91%。此外，我们在永生化的人支气管上皮细胞系16HBE细胞中敲除GSTP1，然后测量它们对致癌物苯并芘诱导的细胞转化的易感性。结果表明，GSTP1敲除显著提高了苯并(a)芘诱导的16HBE细胞转化效率。本研究数据首次表明GSTP1、HSPB1和CKB是LSCC早期检测的新型潜在生物标志物，GSTP1下调参与了人支气管上皮癌的发生。中国生物医学工程学报（自然科学版），2011,32(2):444 - 444。
To discover novel biomarkers for early detection of human lung squamous cell cancer (LSCC) and explore possible mechanisms of LSCC carcinogenesis, iTRAQ-tagging combined with two dimensional liquid chromatography tandem MS analysis was used to identify differentially expressed proteins in human bronchial epithelial carcinogenic process using laser capture microdissection-purified normal bronchial epithelium (NBE), squamous metaplasia (SM), atypical hyperplasia (AH), carcinoma in situ (CIS) and invasive LSCC. As a result, 102 differentially expressed proteins were identified, and three differential proteins (GSTP1, HSPB1 and CKB) showing progressively expressional changes in the carcinogenic process were selectively validated by Western blotting. Immunohistochemistry was performed to detect the expression of the three proteins in an independent set of paraffin-embedded archival specimens including various stage tissues of bronchial epithelial carcinogenesis, and their ability for early detection of LSCC was evaluated by receiver operating characteristic analysis. The results showed that the combination of the three proteins could perfectly discriminate NBE from preneoplastic lesions (SM, AH and CIS) from invasive LSCC, achieving a sensitivity of 96% and a specificity of 92% in discriminating NBE from preneoplatic lesions, a sensitivity of 100% and a specificity of 98% in discriminating NBE from invasive LSCC, and a sensitivity of 92% and a specificity of 91% in discriminating preneoplatic lesions from invasive LSCC, respectively. Furthermore, we knocked down GSTP1 in immortalized human bronchial epithelial cell line 16HBE cells, and then measured their susceptibility to carcinogen benzo(a) pyrene-induced cell transformation. The results showed that GSTP1 knockdown significantly increased the efficiency of benzo(a) pyrene-induced 16HBE cell transformation. The present data first time show that GSTP1, HSPB1 and CKB are novel potential biomarkers for early detection of LSCC, and GSTP1 down-regulation is involved in human bronchial epithelial carcinogenesis. Molecular & Cellular Proteomics 11: 10.1074/mcp.M111.013946, 1-16, 2012.