The mechanisms of highly metastetic capasity in highly metastatic subpopulations of lung adenocarcinoma cell line and these clinical applications

为了寻找新的肺癌肿瘤转移的关键因素，我们建立了一个高转移性的人肺腺癌细胞系作为实验转移模型，通过多次接种裸鼠，随后，培养从肺转移灶中收获的肿瘤细胞，我们利用全基因组人类cDNA微阵列分析建立了两种腺癌细胞系变异的基因表达谱，并将这些谱与亲本细胞系的基因表达谱进行了比较。我们在高转移细胞系中发现了新的基因，这些基因的表达显著增强或减少。我们对肺癌细胞系亚群的分析表明，肺癌的高度转移潜力可能不是由单个基因表达的改变引起的，而是由几个基因表达改变的积累引起的。在本研究中，我们利用蛋白质组分析（2D-DIGE, Ab-array）和新开发的cDNA阵列，鉴定了82个新的候选物，包括半乳糖凝集素-1。我们正在评估这些基因作为参与肺癌转移的关键分子；(1)通过实时RT-PCR、Northern blot和Western blot证实这些表达，(2)通过SiRNA等证实这些功能，(3)通过免疫组织化学在临床标本中评估这些分子的临床应用价值。

To identify novel key factors of tumor metastasis in lung cancer, we established a highly metastatic human lung adenocarcinoma cell line in an experimental metastasis model by repeatedly inoculating the cells into nude mice and, subsequently, culturing tumor cells harvested from the pulmonary metastatic foci and we established the gene expression profiles of two adenocarcinoma cell line variants by use of genome-wide human cDNA microarray analysis and comparing these profiles with that of the parental cell line. We identified novel genes in the highly metastatic cell lines that showed a significantly enhanced or reduced expression. Our analysis in subpopulations of a lung cancer cell line indicated that the highly metastatic potential of lung cancer may be induced not by an alteration in the expression of a single gene, but by the accumulation of alterations in the expression of several genes. In this study, we used proteome analysis (2D-DIGE, Ab-array) and newly-developed cDNA array and identified 82 novel candidates including galectin-1. We are evaluating these genes as key molecules involved in metastasis in lung cancer ; (1)confirm these expressions by real-time RT-PCR, Northern blot, and Western blot, (2)confirm these function by SiRNA et al., (3)evaluate clinical utility of these molecules by immunohistochemistry in clinical specimens.