The promoter of miR-663 is hypermethylated in Chinese pediatric acute myeloid leukemia (AML)

BMC Medical Genetics

Pan Jian²  Feng Xing¹  Wang Jian¹  Pang Li¹  Wu Dong¹  Zhao Wen-Li¹  Xiao Pei-Fang¹  Wang Na¹  Lu Jun¹  Ni Jian²  Tao Yan-Fang¹ 

[1] Department of Hematology and Oncology, Children’s Hospital of Soochow University, Suzhou, China;Translational Research Center, Second Hospital, The Second Clinical School, Nanjing Medical University, Nanjing, China

关键词: Hypermethylation;    MiR-663;    Acute myeloid leukemia;    Pediatric;   

Others  :  1127655
DOI  :  10.1186/1471-2350-14-74

 received in 2012-08-23, accepted in 2013-07-18,  发布年份 2013

【 摘 要 】

Background

There is growing evidence supporting a role for microRNAs (miRNA) as targets in aberrant mechanisms of DNA hypermethylation. Epigenetic silencing of tumor suppressor miRNAs, including miR-663, which has recently been reported to be inactivated by hypermethylation in several cancers, may play important roles in pediatric acute myeloid leukemia (AML). However, expression of miR-663 and its promoter methylation remain status unclear in childhood leukemia.

Methods

Promoter methylation status of miR-663 was investigated by methylation specific PCR (MSP) and bisulfate genomic sequencing (BGS). Transcriptional expression of miR-663 was evaluated by semi-quantitative and real-time PCR, and the relationship between expression of miR-663 and promoter methylation was confirmed using 5-aza-2’-deoxycytidine (5-Aza) demethylation reagent.

Results

MiR-663 was aberrantly methylated in 45.5% (5/11) leukemia cell lines; BGS showed that the promoter was significantly methylated in three AML cell lines; methylation of miR-663 was significantly higher in Chinese pediatric AML patients [41.4% (29/70)] compared to normal bone marrow (NBM) control samples [10.0% (3/30)]. These results were confirmed by both BGS and 5-Aza demethylation analysis. In addition, miR-663 transcript expression was significantly lower in AML patients, both with and without miR-663 methylation, compared to controls; however, there were no significant differences in clinical features or French-American-British (FAB) classification between patients with and without miR-663 methylation.

Conclusions

Expression of miR-663 was significantly lower in pediatric AML cells compared to NBM controls; furthermore, a high frequency of miR-663 promoter hypermethylation was observed in both AML cell lines and pediatric AML samples. Inactivation of miR-663 by promoter hypermethylation could be affected by 5-Aza demethylation. These findings suggest that hypermethylation of the miR-663 promoter may be an early event in the development of pediatric AML.

【 授权许可】

   
2013 Yan-Fang et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Estey E, Dohner H: Acute myeloid leukaemia. Lancet 2006, 368(9550):1894-1907.
• [2]Plass C, Oakes C, Blum W, Marcucci G: Epigenetics in acute myeloid leukemia. Semin Oncol 2008, 35(4):378-387.
• [3]Issa JP: CpG island methylator phenotype in cancer. Nat Rev Cancer 2004, 4(12):988-993.
• [4]Boultwood J, Wainscoat JS: Gene silencing by DNA methylation in haematological malignancies. Br J Haematol 2007, 138(1):3-11.
• [5]Savickiene J, Treigyte G, Borutinskaite VV, Navakauskiene R: Antileukemic activity of combined epigenetic agents, DNMT inhibitors zebularine and RG108 with HDAC inhibitors, against promyelocytic leukemia HL-60 cells. Cell Mol Biol Lett 2012, 17(4):501-525.
• [6]Trowbridge JJ, Sinha AU, Zhu N, Li M, Armstrong SA, Orkin SH: Haploinsufficiency of Dnmt1 impairs leukemia stem cell function through derepression of bivalent chromatin domains. Genes Dev 2012, 26(4):344-349.
• [7]Liu L, Ling X, Liang H, Gao Y, Yang H, Shao J, Tang H: Hypomethylation mediated by decreased DNMTs involves in the activation of proto-oncogene MPL in TK6 cells treated with hydroquinone. Toxicol Lett 2012, 209(3):239-245.
• [8]Walter MJ, Ding L, Shen D, Shao J, Grillot M, McLellan M, Fulton R, Schmidt H, Kalicki-Veizer J, O’Laughlin M, et al.: Recurrent DNMT3A mutations in patients with myelodysplastic syndromes. Leukemia 2011, 25(7):1153-1158.
• [9]Inaba T: [Epidemiology of leukemia and MDS among atomic bomb survivors in Hiroshima and Nagasaki suggests how abnormal epigenetic regulation contributes to leukemogenesis]. Rinsho Ketsueki 2009, 50(10):1548-1552.
• [10]Lehmann U, Hasemeier B, Romermann D, Muller M, Langer F, Kreipe H: Epigenetic inactivation of microRNA genes in mammary carcinoma. Verh Dtsch Ges Pathol 2007, 91:214-220.
• [11]Stumpel DJ, Schotte D, Lange-Turenhout EA, Schneider P, Seslija L, De Menezes RX, Marquez VE, Pieters R, Den Boer ML, Stam RW: Hypermethylation of specific microRNA genes in MLL-rearranged infant acute lymphoblastic leukemia: major matters at a micro scale. Leukemia 2011, 25(3):429-439.
• [12]Svedruzic ZM, Reich NO: DNA cytosine C5 methyltransferase Dnmt1: catalysis-dependent release of allosteric inhibition. Biochemistry 2005, 44(27):9472-9485.
• [13]Garzon R, Liu S, Fabbri M, Liu Z, Heaphy CE, Callegari E, Schwind S, Pang J, Yu J, Muthusamy N, et al.: MicroRNA-29b induces global DNA hypomethylation and tumor suppressor gene reexpression in acute myeloid leukemia by targeting directly DNMT3A and 3B and indirectly DNMT1. Blood 2009, 113(25):6411-6418.
• [14]Afonyushkin T, Oskolkova OV, Bochkov VN: Permissive role of miR-663 in induction of VEGF and activation of the ATF4 branch of unfolded protein response in endothelial cells by oxidized phospholipids. Atherosclerosis 2012, 225(1):50-55.
• [15]Pan J, Hu H, Zhou Z, Sun L, Peng L, Yu L, Sun L, Liu J, Yang Z, Ran Y: Tumor-suppressive mir-663 gene induces mitotic catastrophe growth arrest in human gastric cancer cells. Oncol Rep 2010, 24(1):105-112.
• [16]Pizzimenti S, Ferracin M, Sabbioni S, Toaldo C, Pettazzoni P, Dianzani MU, Negrini M, Barrera G: MicroRNA expression changes during human leukemic HL-60 cell differentiation induced by 4-hydroxynonenal, a product of lipid peroxidation. Free Radic Biol Med 2009, 46(2):282-288.
• [17]Lehmann U, Hasemeier B, Christgen M, Muller M, Romermann D, Langer F, Kreipe H: Epigenetic inactivation of microRNA gene hsa-mir-9-1 in human breast cancer. J Pathol 2008, 214(1):17-24.
• [18]Jian P, Li ZW, Fang TY, Jian W, Zhuan Z, Mei LX, Yan WS, Jian N: Retinoic acid induces HL-60 cell differentiation via the upregulation of miR-663. J Hematol Oncol 2011, 4:20. BioMed Central Full Text
• [19]Tili E, Michaille JJ, Adair B, Alder H, Limagne E, Taccioli C, Ferracin M, Delmas D, Latruffe N, Croce CM: Resveratrol decreases the levels of miR-155 by upregulating miR-663, a microRNA targeting JunB and JunD. Carcinogenesis 2010, 31(9):1561-1566.
• [20]Tili E, Michaille JJ: Resveratrol, MicroRNAs, inflammation, and cancer. J Nucleic Acids 2011, 2011:102431.
• [21]Tili E, Michaille JJ, Alder H, Volinia S, Delmas D, Latruffe N, Croce CM: Resveratrol modulates the levels of microRNAs targeting genes encoding tumor-suppressors and effectors of TGFbeta signaling pathway in SW480 cells. Biochem Pharmacol 2010, 80(12):2057-2065.
• [22]Liu ZY, Zhang GL, Wang MM, Xiong YN, Cui HQ: MicroRNA-663 targets TGFB1 and regulates lung cancer proliferation. Asian Pac J Cancer Prev 2011, 12(11):2819-2823.
• [23]Ni CW, Qiu H, Jo H: MicroRNA-663 upregulated by oscillatory shear stress plays a role in inflammatory response of endothelial cells. Am J Physiol Heart Circ Physiol 2011, 300(5):1762-1769.
• [24]Yi C, Wang Q, Wang L, Huang Y, Li L, Liu L, Zhou X, Xie G, Kang T, Wang H, et al.: MiR-663, a microRNA targeting p21(WAF1/CIP1), promotes the proliferation and tumorigenesis of nasopharyngeal carcinoma. Oncogene 2012, 31(41):4421-4433.
• [25]Cheng Y, Geng H, Cheng SH, Liang P, Bai Y, Li J, Srivastava G, Ng MH, Fukagawa T, Wu X, et al.: KRAB zinc finger protein ZNF382 is a proapoptotic tumor suppressor that represses multiple oncogenes and is commonly silenced in multiple carcinomas. Cancer Res 2010, 70(16):6516-6526.
• [26]Yang Y, Wang LL, Li YH, Gao XN, Liu Y, Yu L: Effect of CpG island methylation on microRNA expression in the k-562 cell line. Biochem Genet 2012, 50(1–2):122-134.
• [27]Yang Y, Wang LL, Li YH, Gao XN, Yu L: [Expression level of miRNA-663 in different leukemic cell lines and its biological function]. Zhongguo Shi Yan Xue Ye Xue Za Zhi 2011, 19(2):279-283.