Identification and validation of metastasis-associated proteins in head and neck cancer cell lines by two-dimensional electrophoresis and mass spectrometry
Identification and validation of metastasis-associated proteins in head and neck cancer cell lines by two-dimensional electrophoresis and mass spectrometry
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DOI：
10.1023/a:1015515119300
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发表时间：
2002-01-01
影响因子：
4
通讯作者：
Mao, L
中科院分区：
文献类型：
--
作者：
Wu, WG;Tang, XM;Mao, L
尽管在过去的二十年中头颈部鳞状细胞癌（HNSCC）患者的治疗有所改善，但这些患者的生存率并没有显著提高。该疾病预后不良的主要因素之一是局部转移。为了在蛋白质水平上更好地了解这一过程的机制，我们使用SELDI ProteinChip技术进行了二维电泳（2-DE）和质谱分析，以鉴定来自同一患者的两种HNSCC细胞系UMSCC10A和UMSCC10B中的差异表达蛋白。UMSCC10A来源于原发肿瘤，UMSCC10B来源于转移性淋巴结。从凝胶中切除差异表达蛋白。在胰蛋白酶凝胶消化后，生成肽的质量谱。通过将肽质量谱提交到公共可用的NCBInr数据库（www.proteometrics.com）来鉴定蛋白质。在转移细胞系UMSCC10B中发现两种膜相关蛋白，膜联蛋白I和膜联蛋白II以及糖酵解蛋白烯醇化酶- α上调，钙蛋白前体下调。通过分析内源性蛋白酶赖氨酸- c酶切获得的肽质量指纹图谱，证实了这些蛋白的身份。Western blotting分析也验证了结果。结果表明，烯醇化酶- α、膜联蛋白i和膜联蛋白ii可能是头颈部肿瘤侵袭转移的重要分子。结果还表明，蛋白质组学在鉴定癌症发生和进展中重要的分子异常方面具有重要的补充作用。
Despite improvements in treatment of patients with head and neck squamous cell carcinoma (HNSCC) over the last two decades, the survival rate of these patients has not increased significantly. One of the major factors in the poor outcome of the disease is regional metastasis. To better understand the mechanisms of this process at the protein level, we performed two-dimensional electrophoresis (2-DE) and mass spectrometry using SELDI ProteinChip technology to identify proteins differentially expressed in two HNSCC cell lines, UMSCC10A and UMSCC10B, from the same patient. UMSCC10A was derived from the primary tumor and UMSCC10B from a metastatic lymph node. The differentially expressed proteins were excised from the gels. Following in-gel digestion by trypsin, mass profiles of the peptides were generated. Proteins were identified by submitting the peptide mass profiles to a public available NCBInr databases (www.proteometrics.com). Two membrane-associated proteins, annexin I and annexin II, and glycolytic protein enolase-alpha were found to be upregulated, and calumenin precursor down-regulated, in metastatic cell line UMSCC10B. The identity of these proteins was confirmed by analyzing additional peptide mass fingerprints obtained by endoproteinase lysine-C digestion. The results were also validated by Western blotting analysis. Our results showed that enolase-alpha, annexin-I and annexin-II might be important molecules in head and neck cancer invasion and metastasis. The results also suggest an important complementary role for proteomics in identification of molecular abnormalities important in cancer development and progression.