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Article: Epigenetic silencing of miR-340-5p in multiple myeloma: mechanisms and prognostic impact

TitleEpigenetic silencing of miR-340-5p in multiple myeloma: mechanisms and prognostic impact
Authors
KeywordsDNA methylation
miR-340-5p
Multiple myeloma
Overall survival
XIAP
Issue Date2019
PublisherBioMed Central Ltd. The Journal's web site is located at http://www.clinicalepigeneticsjournal.com
Citation
Clinical Epigenetics, 2019, v. 11, article no. 71 How to Cite?
AbstractBackground: miR-340-5p, localized to 5q35, is a tumor suppressor miRNA implicated in multiple cancers. As a CpG island is present at the putative promoter region of its host gene, RNF130, we hypothesized that the intronic miR-340-5p is a tumor suppressor miRNA epigenetically silenced by promoter DNA methylation of its host gene in multiple myeloma. Results: By pyrosequencing-confirmed methylation-specific PCR, RNF130/miR-340 was methylated in 8/15 (53.3%) myeloma cell lines but not normal plasma cells. Methylation correlated inversely with the expression of both miR-340-5p and RNF130. In completely methylated WL-2 and RPMI-8226R cells, 5-AzadC treatment led to demethylation and re-expression of miR-340-5p. In primary samples, RNF130/miR-340 methylation was detected in 4 (22.2%) monoclonal gammopathy of undetermined significance, 15 (23.8%) diagnostic myeloma, and 7 (23.3%) relapsed myeloma. RNF130/miR-340 methylation at diagnosis was associated with inferior overall survival (median 27 vs. 68 months; P = 3.944E−5). In WL-2 cells, overexpression of miR-340-5p resulted in reduced cellular proliferation [MTS, P = 0.002; verified in KMS-12-PE (P = 0.002) and RPMI-8226R (P = 2.623E−05) cells], increased cell death (trypan blue, P = 0.005), and enhanced apoptosis by annexin V-PI staining. Moreover, by qRT-PCR, overexpression of miR-340-5p led to repression of both known targets (CCND1 and NRAS) and bioinformatically predicted targets in MAPK signaling (MEKK1, MEKK2, and MEKKK3) and apoptosis (MDM4 and XIAP), hence downregulation of phospho-ERK1/2 and XIAP by Western blot. Furthermore, by qRT-PCR, in CD138-sorted primary samples (n = 37), miR-340-5p and XIAP were inversely correlated (P = 0.002). By luciferase assay, XIAP was confirmed as a direct target of miR-340-5p via targeting of the distal but not proximal seed region binding site. Conclusions: Collectively, tumor-specific methylation-mediated silencing of miR-340-5p is likely an early event in myelomagenesis with adverse survival impact, via targeting multiple oncogenes in MAPK signaling and apoptosis, thereby a tumor suppressive miRNA in myeloma.
Persistent Identifierhttp://hdl.handle.net/10722/272288
ISSN
2019 Impact Factor: 5.028
2015 SCImago Journal Rankings: 2.462
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorLi, Z-
dc.contributor.authorWong, KY-
dc.contributor.authorCalin, GA-
dc.contributor.authorChng, WJ-
dc.contributor.authorChan, GCF-
dc.contributor.authorChim, CS-
dc.date.accessioned2019-07-20T10:39:20Z-
dc.date.available2019-07-20T10:39:20Z-
dc.date.issued2019-
dc.identifier.citationClinical Epigenetics, 2019, v. 11, article no. 71-
dc.identifier.issn1868-7075-
dc.identifier.urihttp://hdl.handle.net/10722/272288-
dc.description.abstractBackground: miR-340-5p, localized to 5q35, is a tumor suppressor miRNA implicated in multiple cancers. As a CpG island is present at the putative promoter region of its host gene, RNF130, we hypothesized that the intronic miR-340-5p is a tumor suppressor miRNA epigenetically silenced by promoter DNA methylation of its host gene in multiple myeloma. Results: By pyrosequencing-confirmed methylation-specific PCR, RNF130/miR-340 was methylated in 8/15 (53.3%) myeloma cell lines but not normal plasma cells. Methylation correlated inversely with the expression of both miR-340-5p and RNF130. In completely methylated WL-2 and RPMI-8226R cells, 5-AzadC treatment led to demethylation and re-expression of miR-340-5p. In primary samples, RNF130/miR-340 methylation was detected in 4 (22.2%) monoclonal gammopathy of undetermined significance, 15 (23.8%) diagnostic myeloma, and 7 (23.3%) relapsed myeloma. RNF130/miR-340 methylation at diagnosis was associated with inferior overall survival (median 27 vs. 68 months; P = 3.944E−5). In WL-2 cells, overexpression of miR-340-5p resulted in reduced cellular proliferation [MTS, P = 0.002; verified in KMS-12-PE (P = 0.002) and RPMI-8226R (P = 2.623E−05) cells], increased cell death (trypan blue, P = 0.005), and enhanced apoptosis by annexin V-PI staining. Moreover, by qRT-PCR, overexpression of miR-340-5p led to repression of both known targets (CCND1 and NRAS) and bioinformatically predicted targets in MAPK signaling (MEKK1, MEKK2, and MEKKK3) and apoptosis (MDM4 and XIAP), hence downregulation of phospho-ERK1/2 and XIAP by Western blot. Furthermore, by qRT-PCR, in CD138-sorted primary samples (n = 37), miR-340-5p and XIAP were inversely correlated (P = 0.002). By luciferase assay, XIAP was confirmed as a direct target of miR-340-5p via targeting of the distal but not proximal seed region binding site. Conclusions: Collectively, tumor-specific methylation-mediated silencing of miR-340-5p is likely an early event in myelomagenesis with adverse survival impact, via targeting multiple oncogenes in MAPK signaling and apoptosis, thereby a tumor suppressive miRNA in myeloma.-
dc.languageeng-
dc.publisherBioMed Central Ltd. The Journal's web site is located at http://www.clinicalepigeneticsjournal.com-
dc.relation.ispartofClinical Epigenetics-
dc.rightsThis work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.-
dc.subjectDNA methylation-
dc.subjectmiR-340-5p-
dc.subjectMultiple myeloma-
dc.subjectOverall survival-
dc.subjectXIAP-
dc.titleEpigenetic silencing of miR-340-5p in multiple myeloma: mechanisms and prognostic impact-
dc.typeArticle-
dc.identifier.emailWong, KY: kwanumu@hku.hk-
dc.identifier.emailChan, GCF: gcfchan@hku.hk-
dc.identifier.emailChim, CS: jcschim@hku.hk-
dc.identifier.authorityChan, GCF=rp00431-
dc.identifier.authorityChim, CS=rp00408-
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.1186/s13148-019-0669-2-
dc.identifier.pmid31064412-
dc.identifier.pmcidPMC6505104-
dc.identifier.scopuseid_2-s2.0-85065503631-
dc.identifier.hkuros299540-
dc.identifier.volume11-
dc.identifier.spagearticle no. 71-
dc.identifier.epagearticle no. 71-
dc.identifier.isiWOS:000467444000001-
dc.publisher.placeUnited Kingdom-

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