背景与目的肿瘤转移抑制基因nm23-H1基因调控肺癌细胞转移潜能的确切机理尚未明了,本研究旨在探讨人高转移大细胞肺癌细胞株(L9981)和转染nm23-H1基因的人高转移大细胞肺癌细胞株(L9981-nm23-H1)的差异表达蛋白,为阐明肺癌转移的分子机制、发现早期诊断肺癌转移的分子标志和新的治疗靶点提供实验依据。方法应用固相pH梯度双向凝胶电泳分离L9981和L9981-nm23-H1细胞株的总蛋白,对25个差异明显的蛋白质点进行质谱鉴定和生物信息学分析。结果研究观察到nm23-H1基因转染使人L9981细胞株蛋白质组的表达谱发生了明显变化:5个蛋白质表达缺失,9个新的蛋白表达,16个蛋白质表达下调,12个蛋白质表达上调。这些蛋白质主要涉及细胞骨架蛋白、信号转导蛋白、细胞代谢相关蛋白、发育增殖相关蛋白及肿瘤侵袭相关蛋白。结论 nm23-H1基因转染L9981后,蛋白质表达谱发生了显著的变化,这些差异蛋白质可能是逆转肺癌侵袭转移的生化基础,本研究结果可能为阐明肺癌转移的分子机制提供线索。
Background and objective As a tumor metastasis suppressor gene, the functions of nm23-H1 gene are still unclear. The aim of this study is to better understand the mechanism of lung cancer metastasis and to find new biomark-ers for early diagnosis and new target for therapy by conducting comparative proteomics between the human high-metastatic large cell lung cancer cell lines (L9981) and L9981-nm23-H1 (constructed with transfecting nm23-H1 gene into the L9981 cell line). Methods The total proteins of L9981 and L9981-nm23-H1 were separated by immobilized pH gradient (IPG)-based 2-dimensional electrophoresis (2-DE); the significantly differently expressed proteins were examined by mass spectrometry and analyzed by bioinformatics. Results It was observed that nm23-H1 gene transfection caused remarkable changes of the proteome of L9981 compared with L9981-nm23-H1 cells: 5 proteins were deleted, 9 proteins appeared, 16 proteins down-regulated, and 12 proteins up-regulated. These proteins are involved in cell framework, signal transduction, metabolism, proliferation and metastasis. Conclusion After nm23-H1 gene is transfected into L9981, proteome in L9981 is remarkably changed. These changes of the proteome could serve as a basis for reversing the invasive and metastatic phenotype in lung cancer and elucidating the machanisms of the metastasis of lung cancer.