辛伐他汀对人慢性粒细胞白血病细胞K562基因表达的影响

Effect of simvastatin on gene expression profiles of human chronic myelocytic leukemia cell line K562

目的应用基因芯片技术研究辛伐他汀对K562细胞基因表达的影响,初步探讨其作用机理。方法辛伐他汀作用K562细胞48h后,提取细胞总RNA。纯化mRNA后再逆转录为cDNA,将cDNA在体外转录合成cRNA,用随机引物反转录为cDNA,用Klenow酶标记Cy3、Cy5,与定制的包含人全基因组的基因芯片杂交,扫描芯片荧光信号,用GenePixPro4.0图像分析软件对芯片图像进行分析。定量RT-PCR验证芯片结果中两个差异表达基因。结果基因芯片检测发现共有176条基因表达发生明显改变,包括16条未知基因表达改变。其中151条基因表达上调,25条基因表达下调。根据基因功能的不同,我们初步将其分为凋亡相关、信号转导相关、细胞周期相关等几大类。定量RT-PCR验证的基因表达变化趋势与表达谱芯片的结果相吻合。结论辛伐他汀作用K562细胞后能引起多种基因表达改变,这为探讨其作用机制提供了新的依据。

Objective To investigate the effect of simvastatin on gene expression＇profiles of K562 cells and to elucidate its mechanism. Methods The total RNA of K562 cells treated with or without simvastatin for 48 h was extracted and purified, followed by cDNA synthesis, then in vitro transcription reaction was subjected to synthesizing cRNA, which was transcripted into cDNA using random primers, cDNA labelled Cy3 and Cy5 by Klenow enzyme was hybridized with genechip, then the chip was scanned with ScanArray Express Scanner, and obtained images were analyzed with GenePix Pro4.0. Finally, two differentially regulated genes were validated by real time RT-PCR. Results There were 176 differential expression genes between simvastatin-treated group and untreated group, including 16 unidentified genes. Expression of 25 genes was down-regulated and that of 151 genes was up-regulated in the treatment group compared to the untreated group. According to the functions of these genes, they were classified into apoptosis-related, signal transduction and cell cycle related genes. Two differential expression genes in genechip were conformed by real time RT-PCR. Conclusion Simvastatin can regulate expression of many genes in K562 cells, which might provide novel foundation for its anti-cancer mechanism.