nm23-H1基因转染L9981肺癌细胞前后基因表达谱的变化

Analysis of the Gene Expression Changes in Human High-metastatic Large Cell Lung Cancer Cell Line L9981 Betected by Microarray Before and After Transfection with nm23-H1 Gene

应用基因芯片检测L9981细胞转染nm23-H1基因前后细胞基因表达谱的改变.提取L9981细胞转染nm23-H1基因前后细胞的总RNA,纯化为mRNA后再转录为cDNA.cDNA经限制性内切酶Sau3AI消化后,cDNA片段分别用cy3和cy5标记,与定制的包含14000个基因芯片杂交.杂交结果经扫描和软件分析,nm23-H1基因转染L9981细胞后发现1156(8.26%,1156/14000)个基因表达上调,而642(4.59%,642/14000)个基因表达下调.涉及基因包括信号传导、癌基因与抑癌基因、转移相关基因、细胞周期与凋亡、细胞外基质与细胞骨架相关基因,以及细胞因子和转录因子等.nm23-H1基因是通过对转移相关基因的调节来发挥其抑制肺癌细胞株L9981侵袭和转移作用的.

To investigate the changes of gene expression in the human high-metastatic large cell lung cancer cell line L9981 transfected and untransfected with nm23-H1 gene. The L9981 and L9981-nm23-H1 cells were cuhured in RPMI 1 640 medium supplemented with 10% calf serum. Total RNA of the L9981 and L9981-nm23-H1 cells were then extracted, purified and subject to reverse transcription into first-strand cDNA, labeled with Cy3/Cy5. Placenta DNA microarrays containing 14 000 DNA fragments were used to analyze the differential gene expression of L9981 and L9981-nm23-H1 cells. 1 156 genes （8.26%, 1 156/14 000） were upregulated and 642 genes （4.59%, 642/14 000） were downregulated in the nm23 transfected cell line, including many different types of genes, such as those responsible for signal transduction, oncogene and tumor suppression genes, metastatic-associated genes, apoptatic-related genes and DNA binding and transcription factor genes. These data suggested that the molecular mechanism by which nm23-H1 suppresses L9981 cells invasion and metastasis may be via regulating the expression of metastatic-related genes.