Determination of metastasis-associated proteins in non-small cell lung cancer by comparative proteomic analysis
Determination of metastasis-associated proteins in non-small cell lung cancer by comparative proteomic analysis
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DOI：
10.1111/j.1349-7006.2007.00514.x
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发表时间：
2007-08-01
期刊：
影响因子：
5.7
通讯作者：
He, Dacheng
中科院分区：
文献类型：
--
作者：
Tian, Tian;Hao, Jia;He, Dacheng
在非小细胞肺癌的病例中，转移的发展是死亡的主要原因和巨大的治疗挑战。为了更好地理解转移过程的分子机制并发现非小细胞肺癌的新的潜在临床标志物，对具有不同转移潜能的两种非小细胞肺癌细胞系（非转移性CL 1 -0和高转移性CL 1 -5细胞系）进行比较蛋白质组学分析，进行了使用二维电泳，然后基质辅助激光解吸电离飞行时间质谱和串联质谱。共鉴定出33个差异表达蛋白，其中16个蛋白在高转移性CL 1 -5细胞中表达上调，17个蛋白表达下调。随后，从33个鉴定的蛋白质中选择8个，使用实时定量聚合酶链反应在mRNA水平上进行进一步验证，并通过蛋白质印迹法确认了3个鉴定的蛋白质，S100 A11，PGP 9.5和HSP 27。对65例原发性非小细胞肺癌组织和10例配对的局部阳性淋巴结标本进行S100 A11蛋白免疫组化染色，分析S100 A11蛋白在蛋白和mRNA水平的差异表达，探讨其与转移的关系。结果表明，S100 A11在非小细胞肺癌组织中的表达与肿瘤的分期、淋巴结转移密切相关（P = 0.001），与淋巴结转移密切相关（P = 0.011），提示S100 A11可能是促进非小细胞肺癌侵袭转移的重要调控分子。
The development of metastasis is the leading cause of death and an enormous therapeutic challenge in cases of non-small cell lung cancer. To better understand the molecular mechanisms underlying the metastasis process and to discover novel potential clinical markers for non-small cell lung cancer, comparative proteomic analysis of two non-small cell lung cancer cell lines with different metastatic potentials, the non-metastatic CL1-0 and highly metastatic CL1-5 cell lines, was carried out using two-dimensional electrophoresis followed by matrix-assisted laser desorption ionization-time of flight mass spectrometry and tandem mass spectrometry. Thirty-three differentially expressed proteins were identified unambiguously, among which 16 proteins were significantly upregulated and 17 proteins were downregulated in highly metastatic CL1-5 cells compared with non-metastatic CL1-0 cells. Subsequently, 8 of 33 identified proteins were selected for further validation at the mRNA level using real-time quantitative polymerase chain reaction, and three identified proteins, S100A11, PGP 9.5 and HSP27, were confirmed by western blotting. The protein S100A11 displaying significant differential expression at both the protein and mRNA levels was further analyzed by immunohistochemical staining in 65 primary non-small cell lung cancer tissues and 10 matched local positive lymph node specimens to explore its relationship with metastasis. The results indicated that the upregulation of S100A11 expression in non-small cell lung cancer tissues was significantly associated with higher tumor-node-metastasis stage (P = 0.001) and positive lymph node status (P = 0.011), implying that S100A11 might be an important regulatory molecule in promoting invasion and metastasis of non-small cell lung cancer.