采用人基因表达谱芯片筛选肺癌转移相关基因的初步研究  

Screening of metastasis-associated genes in human lung cancer using cDNA microarray chip

作　　者：常艳[1] 陈良安[1] 田庆[1] 王平[1] 梁志欣[1] 范保星[1] 

机构地区：[1]解放军总医院呼吸科,北京市100853

出　　处：《中华老年多器官疾病杂志》2007年第1期47-51,58,共6页Chinese Journal of Multiple Organ Diseases in the Elderly

基　　金：国家自然科学基金资助课题(30370616)

摘　　要：目的应用基因芯片技术分析人高、低转移肺巨细胞癌细胞株的基因差异表达谱,筛选与肺癌转移相关基因。方法提取人高转移肺巨细胞株95D和人低转移肺巨细胞株95C的mRNA,通过逆转录,制备分别用cy5-dUTP和cy3-dUTP进行标记的cDNA探针,并与芯片杂交。经过ScanArray3000扫描仪扫描,获取图象及对杂交信号进行数据分析,筛选出95D和95C细胞表达差异的基因谱,并对其中部分基因进行RT-PCR分析、验证。结果在所检测95C和95D细胞中,466条基因有表达差异,其中具有同一Genebank量的有108对,上调基因47对,下调基因61对。对其中KIAA1108、PGR1、JWA、S182、Jab1部分差异表达基因,经RT-PCR技术验证,结果与基因芯片基本符合。结论肺癌转移过程与多种基因作用有关,基因芯片技术可为肺癌转移相关基因的筛选,提供有效方法。Objective To screen lung cancer metastasis-associated genes by analysis of differentially expressed genes between human lung giant cell carcinoma cell strains of high metastatic 95D and low metastatic 95C using cDNA microarray. Methods The mRNAs were extracted from human lung giant cell carcinoma cell strains of 95D and 95C, and reversely transcribed to the cDNAs , which were then labeled by Cy5-dUTP and Cy3-dUTP to prepare the hybridization probes. The mixed probes were hybridized with the cDNA microarray. The figure was obtained by scanning the cDNA microarray chip with Scan Array 3000 scanner and the information was managed with ImaGene3.0 software. Thus the genes chip of different expression between high metastatic 95D and low metastatic 95C was screened. A part of genes whose expression was changed in cDNA microarray analysis were further identified by RTPCR. Results The cDNA microarray analysis showed that the expression of 466 genes was changed . Among them, 108 pairs of double gene had the same Genbank ID, including 47 pairs of up-regulation genes and 61 pairs of down-regulation genes. Fifty-eight genes were not elsewhere classified. Some of the genes, including KIAA1108, PGR1, JWA, S182, Jabl etc, were further confirmed by RT-PCR. The results of RT-PCR coincided well with the cDNA microarray results. Conclusion Many different genes are involved in the metastasis of lung cancer, cDNA microarray technique might be effective in screening lung cancer metastasis-associated genes.

关 键 词：肺肿瘤 基因筛选 肿瘤转移 逆转录-聚合酶链反应 

分 类 号：R735.2[医药卫生—肿瘤]