Abstract
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.
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Acknowledgments
This research was supported by grants of the National Natural Science Foundation of China (Nos. 90919044 and 30971297) and Major State Basic Research Development Program (973 Program No. 2005CB522408). We greatly thank Prof. Xiao-Fei Zheng and Assoc. Prof. Han-Jiang Fu of the Chinese PLA Academy of Military Medical Science for technical assistance in the detection of mature miRNAs.
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Yang, Y., Wang, LL., Li, YH. et al. Effect of CpG Island Methylation on MicroRNA Expression in the k-562 Cell Line. Biochem Genet 50, 122–134 (2012). https://doi.org/10.1007/s10528-011-9478-9
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DOI: https://doi.org/10.1007/s10528-011-9478-9