Analysis of triptolide-regulated gene expression in Jurkat cells by complementary DNA microarray.
Analysis of triptolide-regulated gene expression in Jurkat cells by complementary DNA microarray.
复制标题
通过互补 DNA 微阵列分析 Jurkat 细胞中雷公藤甲素调节的基因表达。
DOI：
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发表时间：
2003
影响因子：
8.2
通讯作者：
Shunyou Wang
中科院分区：
文献类型：
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作者：
Z. Du;Xiao;Yuan;Shunyou Wang
目的
研究雷公藤甲素作用后Jurkat细胞基因表达谱的变化，寻找雷公藤甲素作用的可能靶点。
方法
Jurkat细胞用或不用雷公藤甲素10 μ g/L处理2 h。分离总RNA并用作荧光cDNA探针逆转录标记的模板。使用具有一组13，872个人类基因/EST的高密度DNA微阵列芯片，通过与荧光标记探针杂交来产生雷公藤甲素处理或未处理的对照Jurkat细胞的表达谱。采用基因芯片分析软件GeneSpring对芯片图像进行分析。
结果
雷公藤内酯醇显著抑制Jurkat细胞中117个基因的表达。在这117个基因中，30%是EST或功能未知的基因，13%是转录因子，9%是信号转导通路调节因子，9%是DNA结合蛋白。值得注意的是，丝裂原活化蛋白激酶5（MAP激酶5）和磷酸肌醇-3-激酶（PI-3激酶）的表达被抑制超过100倍。雷公藤内酯醇还下调了脂质转运和代谢相关基因的表达。
结论
高密度微阵列技术为药物靶向分子的筛选提供了一种有效的方法。这表明雷公藤内酯醇的免疫抑制和抗肿瘤作用至少部分是通过抑制MAP激酶和PI-3激酶基因表达介导的。
AIM
To investigate the global gene expression profile changes in Jurkat cells after triptolide treatment in order to find the possible triptolide targets.
METHODS
Jurkat cells were treated with or without triptolide 10 microg/L for 2 h. Total RNA were isolated and used as templates for reverse transcriptional labeling of fluorescent cDNA probes. High density DNA microarray chips with a set of 13,872 human genes/Ests were used to generate the expression profile of triptolide-treated or untreated control Jurkat cells by hybridizing with fluorescent labeled probes. Array image was acquired and analyzed with array analyzing software GeneSpring.
RESULTS
Triptolide significantly suppressed expression of 117 genes in Jurkat cells. Among these 117 genes, 30 % were Ests or genes without known functions, 13 % were transcription factors, 9 % were signal transduction pathway regulators, and 9 % were DNA binding proteins. Notably, the expression of mitogen-activated protein kinase kinase kinase kinase 5 (MAP kinase 5) and phosphoinositide-3-kinase (PI-3 kinase) was inhibited more than 100-fold. Moreover, the expression of genes involved in lipid transportation and metabolism was down-regulated by triptolide.
CONCLUSION
High-density microarray provided an effective approach to identify drug targeting molecules. It is suggested that the widely known immune suppressive and antitumor effects of triptolide were mediated at least in part by suppression of MAP kinase and PI-3 kinase gene expression.