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Effect of CpG island methylation on MicroRNA expression in the k-562 cell line

机译：CpG岛甲基化对k-562细胞系MicroRNA表达的影响

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摘要

To test the hypothesis that methylation of a CpG island is associated with regulation of microRNA expression, we investigated CpG islands in the upstream sequences of microRNA precursors (pre-miRNAs) through bioinformatic analysis and determined whether the CpG islands were methylated by methylation-specific PCR in the k-562 cell line. We used 5-azacytidine for DNA demethylation, and changes in microRNA expression were detected by microarray assay, RT-PCR, and real-time PCR after 5-azacytidine induction. We showed that the CpG islands in the upstream regions of 18 pre-miRNAs were methylated, including miR-663, miR-369, miR-615, and miR-410, and promoter activity was detected in the upstream region of pre-miR-663. We found that a decrease in methylation of a CpG island could up-regulate the expression of miR-663, suggesting that miR-663 could be regulated by DNA methylation. Expression levels of miR-369, miR-615, and miR-410 were not regulated by DNA methylation in this cell line.
机译：为了检验CpG岛甲基化与microRNA表达调控相关的假设，我们通过生物信息学分析研究了microRNA前体（pre-miRNA）上游序列中的CpG岛，并确定甲基化特异性PCR是否将CpG岛甲基化在k-562细胞系中我们使用5-氮杂胞苷进行DNA去甲基化，并通过微阵列测定，RT-PCR和实时PCR检测5-氮杂胞苷诱导后microRNA表达的变化。我们发现18个pre-miRNA上游区域的CpG岛被甲基化，包括miR-663，miR-369，miR-615和miR-410，并且在pre-miR-上游区域检测到启动子活性。 663。我们发现，CpG岛甲基化的减少可以上调miR-663的表达，这表明miR-663可以受到DNA甲基化的调节。在该细胞系中，miR-369，miR-615和miR-410的表达水平不受DNA甲基化的调节。

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