基因表达谱芯片分析人肺腺癌干细胞与肺正常干细胞的差异基因表达  

Analysis of the different gene expression profiles between lung adenocarcinoma stem cells and normal lung stem cells with cDNA microarrays

作　　者：罗福康[1] 赵振国[1] 赵伟鹏[1] 陈熙[1] 陈红[1] 刁鑫伟[1] 谢启超[1] 朱波[1] 孙建国[1] 陈正堂[1] 

机构地区：[1]第三军医大学新桥医院全军肿瘤研究所,重庆400037

出　　处：《中国输血杂志》2009年第4期269-274,共6页Chinese Journal of Blood Transfusion

基　　金：国家高新技术计划(863)资助课题(编号:2007AA02Z129)

摘　　要：目的比较肺腺癌干细胞与正常肺干细胞基因表达水平的差异。方法应用cDNA微阵列技术对肺腺癌和正常肺来源的干细胞的基因表达情况进行检测和比较,采用流式细胞分离技术分离2种干细胞,抽提细胞总RNA、扩增、Cy3染色后,与Agilent 4×44K人全基因组芯片杂交,扫描荧光信号,以Agilent feature extraction数据分析软件分析荧光信号,挑选差异表达基因,进一步对差异表达基因进行GO分析和pathway分析;以qRT-PCR验证芯片结果。结果在肺干细胞全基因组约41 000个基因中,肺腺癌和正常肺来源干细胞的差异表达基因多达5798个(Cut off=2.0);Agilent feature extraction数据分析软件显示2种细胞有明显不同的表达谱;GO分析表明在生物过程中,生物调节相关的差异表达基因占41%,其它比例较高的有细胞组分形成和发生、包内信号级联放大、细胞分化、细胞周期等;在分子功能中,72%的差异表达基因与结合有关,其它比例较高的包括转移酶活性、激酶活性和酶调节活性;在细胞组分方面,分布比较平均,包膜占13%,其它为非膜结合细胞器、胞外区、细胞组分、大分子复合物组分等;pathway分析显示wnt通路有明显差异,共有14个基因表达差异明显。原癌基因中共有11个原癌基因的表达发生了差异性改变,有3个基因上调倍数超过了10,分别是RAB25、MERTK、EGFR,而AKT3下调达到10倍。qRT-PCR结果与芯片结果比较,差异无统计学意义(P>0.05)。结论肺腺癌干细胞与正常肺干细胞在基因表达水平有明显差异。Objective To compare differential gene expression profiles between lung edenocareinoma stem cells and normal lung stem ceils. Methods Lung adenocareinoma and normal lung stem cells were obtained by FACS sorting, and the total RNA was isolated from the cells with Trizol, then the RNA was amplified and dyed with Cy3. The fluorescence was detected and analyzed with Agilent feature extraction; the differentially expressed genes were selected （fold changes over 2. 0 as the cut-offvalue） and GO analysis was performed as well as pathway analysis, qRT-PCR was performed to confirme the microarray results. Results Among 41 000 genes examined, 5 798 genes were differently expressed by the stem cells derived from lung adenocarcinoma and normal lung. Agilent feature extraction showed obviously different expression profiles between the two stem cells. Gene Ontology （GO） analysis showed that 41 percent differentially expressed genes were related to biological regulation within Biological process, and cellular component organization and biogenesis, intracellular signaling cascade, ceil differentiation, and cell cycle were also as a portion of Biological process. In Molecular function, 72 percent differentially expressed genes grouped to binding, and others such as transferase activity, kinase activity, and enzyme regulator activity were commonly represented. Within Cellular component, membrane was 13 percent of this term, and others including nonembrane-bounded organelle, extracellular region, cell fraction, and macromolecular complex assembly distributed evenly. There were 14 differentially expressed genes associated with writ signal pathway and 11 grouped to oncogenes. The results of qRT-PCR and microarrays showed no significant difference （ P 〉 0. 05 ）. Conclusion Our findings suggest that patters of gene expression in the lung adenocarcinoma stem cell and normal lung stem cell are obviously different.

关 键 词：肺 肺腺癌 干细胞 基因表达 CDNA芯片 统计学分析 基因本体论 信号通路 

分 类 号：R446.1[医药卫生—诊断学] R734.2[医药卫生—临床医学]