Gao Liwei, Zhu Wen, Li Lu, Hou Mei, Ma Li, Zhao Ying, Zhou Qinghua
Tianjin Lung Cancer Institute,Tianjin Medical University General Hospital,Tianjin 300052, China.
Zhongguo Fei Ai Za Zhi. 2010 Oct;13(10):928-32. doi: 10.3779/j.issn.1009-3419.2010.10.01.
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 biomarkers 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).
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.
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 downregulated, and 12 proteins up-regulated. These proteins are involved in cell framework, signal transduction, metabolism, proliferation and metastasis.
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 mechanisms of the metastasis of lung cancer.
作为一种肿瘤转移抑制基因,nm23-H1基因的功能仍不清楚。本研究的目的是通过对人高转移性大细胞肺癌细胞系(L9981)和L9981-nm23-H1(通过将nm23-H1基因转染到L9981细胞系构建而成)进行比较蛋白质组学研究,以更好地理解肺癌转移机制,并寻找早期诊断的新生物标志物和新的治疗靶点。
采用基于固定化pH梯度(IPG)的二维电泳(2-DE)分离L9981和L9981-nm23-H1的总蛋白;通过质谱检测差异显著的表达蛋白,并进行生物信息学分析。
观察到与L9981-nm23-H1细胞相比,nm23-H1基因转染导致L9981细胞蛋白质组发生显著变化:5种蛋白缺失,9种蛋白出现,16种蛋白下调,12种蛋白上调。这些蛋白涉及细胞骨架、信号转导、代谢、增殖和转移。
将nm23-H1基因转染到L9981后,L9981中的蛋白质组发生显著变化。这些蛋白质组变化可为逆转肺癌侵袭和转移表型及阐明肺癌转移机制提供依据。