表达谱芯片技术在药物作用机制研究中的应用  

The application of cDNA microarrays technology in mechanism of drug

作　　者：邹明瑾[1] 郭文静[2] 杨美香[3] 张丹[4] 张静[4] 闫实[3] 王景芝[3] 邵倩倩[3] 曲迅[3] 

机构地区：[1]山东大学齐鲁医院检验科,济南250012 [2]山东大学第二附属医院妇产科 [3]山东大学齐鲁医院基础研究所,济南250012 [4]山东中医药大学中西医结合实验室

出　　处：《临床检验杂志》2005年第6期425-428,共4页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金项目(No.30472261)

摘　　要：目的探讨表达谱芯片技术在研究药物作用机制中的价值。方法建立荷NuTu-19卵巢癌的大鼠模型及相应对照,用罗勒多糖治疗50天,处死大鼠,以腹水量、腹腔脏器转移程度积分判断各组肿瘤生长及转移情况。提取癌组织总RNA,用RT-PCR逆转录合成cDNA,分别采用Cy3-dUTP、Cy5-dUTP标记对照组和药物组cDNA,与含有2 000点的大鼠基因表达谱芯片杂交,杂交芯片扫描结果用Im aGene 3.0软件分析,计算Cy3、Cy5两种荧光信号的强度比值。结果对照组与药物组之间共有168条差异表达基因,表达差异在3倍以上的基因有65条,均为表达下调基因;表达差异在2倍以上的基因有103条,其中表达上调基因43条,下调基因60条。经归类分析,表达差异基因主要为:①肿瘤转移相关基因;②与免疫应答有关的基因;③与细胞能量代谢有关的酶类;④与药物代谢及敏感性有关酶类的基因;⑤与信号转导以及转录过程有关的基因等。结论罗勒多糖可能通过抑制肿瘤代谢、调节肿瘤转移相关基因的表达等多种作用途径抑制肿瘤的转移,表达谱芯片技术可以快速确定药物作用的可能机制,并为进一步研究提供重要信息,加快药物的研发速度。Objective To study the application of cDNA microarrays in the mechanism analysis for drug. Methods Rat model bearing NuTu-19 Cells was made and the changes of biology phenotype in tumor cells were observed after treatment with Basil polysaccharide for 50 days. Total RNA of the tumor tissue was extracted, and the fluorescent eDNA probes were prepared through reverse transcription of the isolated mRNAs. The RNA samples were labeled with Cy3-dUTP or Cy5-dUTP, then hybridized with the microarrays. The chips were then scanned with a ScanArray. The acquired images were analyzed using ImaGene 3.0 software. Results Compared with the controls, 168 genes showed different expressions. Sixty-five genes showed more than three times of expression difference, all of which were downregulated. One hundred and three genes showed twice of expression difference, in which 43 genes were upregulated while 60 genes were downregulated. Conclusions eDNA microarray technology may he a powerful tool in discovering the possible mechanism of drug in gene level, provide new informations, and accelerates the development of new drugs.

关 键 词：基因芯片 中草药 罗勒多糖 卵巢癌 

分 类 号：R73[医药卫生—肿瘤]