Methylation-associated Silencing of microRNA-126 and its Host Gene EGFL7 in Malignant Pleural Mesothelioma

Morten Andersen; Davide Trapani; Jesper Ravn; Jens Benn Sørensen; Claus Bøgelund Andersen; Morten Grauslund; Eric Santoni-Rugiu

Methylation-associated Silencing of microRNA-126 and its Host Gene EGFL7 in Malignant Pleural Mesothelioma

Morten Andersen, Davide Trapani, Jesper Ravn, Jens Benn Sørensen, Claus Bøgelund Andersen, Morten Grauslund, Eric Santoni-Rugiu

Department of Clinical Medicine

25 Citations (Scopus)

Abstract

Background/Aim: We recently reported that miR-126 is down-regulated in malignant pleural mesothelioma (MPM) and can be combined into a 4-microRNA-classifier that can accurately diagnose MPM with high sensitivity and specificity. Herein we analyzed the epigenetic regulation of miR-126 and its host gene EGF-like domain, multiple 7 (EGFL7). Materials and Methods: Resected formalin-fixed paraffin-embedded MPM tissues from 29 patients, 14 patientmatched non-neoplastic pleura (NNP) specimens, 5 MPM diagnostic biopsies (DB), and 5 samples of pneumothoraxinduced benign reactive mesothelial proliferation (PTHX) were analyzed. miR-126 and EGFL7 mRNA were quantified by RT-qPCR. CpG-islands' methylation in the EGFL7 promoter was analyzed using methylation-specific PCR and in the MIR126-containing intron 7 was quantified by pyrosequencing. Results: Relative to NNP, EGFL7 was under-expressed more than 4-fold in MPM (p<0.001). EGFL7 mRNA and miR-126 levels correlated in MPM (p<0.01) and NNP (p<0.001). The EGFL7 promoter region was hypermethylated in 69% of MPM and 80% of DB samples, but not in NNP and PTHX samples. EGFL7 promoter hypermethylation was associated with epithelioid histology (p<0.05) and reduced patient-survival (p<0.05). Conclusion: In MPM, DNA-hypermethylation down-regulates miR-126 and its host gene EGFL7, therefore is a poor prognostic factor, and may represent a future therapeutic target for de-methylating strategies re-establishing EGFL7 and miR-126 expression.

Original language	English
Journal	Anticancer Research
Volume	35
Issue number	11
Pages (from-to)	6223-9
Number of pages	7
ISSN	0250-7005
Publication status	Published - Nov 2015

Keywords

Adult
Aged
Biomarkers, Tumor
Case-Control Studies
DNA Methylation
Endothelial Growth Factors
Epigenesis, Genetic
Female
Follow-Up Studies
Gene Expression Regulation, Neoplastic
Humans
Lung Neoplasms
Male
Mesothelioma
MicroRNAs
Middle Aged
Neoplasm Staging
Pleural Neoplasms
Prognosis
Promoter Regions, Genetic
Real-Time Polymerase Chain Reaction

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@article{6b20604907bd4500b086819106f94bb6,

title = "Methylation-associated Silencing of microRNA-126 and its Host Gene EGFL7 in Malignant Pleural Mesothelioma",

abstract = "Background/Aim: We recently reported that miR-126 is down-regulated in malignant pleural mesothelioma (MPM) and can be combined into a 4-microRNA-classifier that can accurately diagnose MPM with high sensitivity and specificity. Herein we analyzed the epigenetic regulation of miR-126 and its host gene EGF-like domain, multiple 7 (EGFL7). Materials and Methods: Resected formalin-fixed paraffin-embedded MPM tissues from 29 patients, 14 patientmatched non-neoplastic pleura (NNP) specimens, 5 MPM diagnostic biopsies (DB), and 5 samples of pneumothoraxinduced benign reactive mesothelial proliferation (PTHX) were analyzed. miR-126 and EGFL7 mRNA were quantified by RT-qPCR. CpG-islands' methylation in the EGFL7 promoter was analyzed using methylation-specific PCR and in the MIR126-containing intron 7 was quantified by pyrosequencing. Results: Relative to NNP, EGFL7 was under-expressed more than 4-fold in MPM (p<0.001). EGFL7 mRNA and miR-126 levels correlated in MPM (p<0.01) and NNP (p<0.001). The EGFL7 promoter region was hypermethylated in 69% of MPM and 80% of DB samples, but not in NNP and PTHX samples. EGFL7 promoter hypermethylation was associated with epithelioid histology (p<0.05) and reduced patient-survival (p<0.05). Conclusion: In MPM, DNA-hypermethylation down-regulates miR-126 and its host gene EGFL7, therefore is a poor prognostic factor, and may represent a future therapeutic target for de-methylating strategies re-establishing EGFL7 and miR-126 expression.",

keywords = "Adult, Aged, Biomarkers, Tumor, Case-Control Studies, DNA Methylation, Endothelial Growth Factors, Epigenesis, Genetic, Female, Follow-Up Studies, Gene Expression Regulation, Neoplastic, Humans, Lung Neoplasms, Male, Mesothelioma, MicroRNAs, Middle Aged, Neoplasm Staging, Pleural Neoplasms, Prognosis, Promoter Regions, Genetic, Real-Time Polymerase Chain Reaction",

author = "Morten Andersen and Davide Trapani and Jesper Ravn and S{\o}rensen, {Jens Benn} and Andersen, {Claus B{\o}gelund} and Morten Grauslund and Eric Santoni-Rugiu",

year = "2015",

month = nov,

language = "English",

volume = "35",

pages = "6223--9",

journal = "Anticancer Research",

issn = "0250-7005",

publisher = "International Institute of Anticancer Research",

number = "11",

}

TY - JOUR

T1 - Methylation-associated Silencing of microRNA-126 and its Host Gene EGFL7 in Malignant Pleural Mesothelioma

AU - Andersen, Morten

AU - Trapani, Davide

AU - Ravn, Jesper

AU - Sørensen, Jens Benn

AU - Andersen, Claus Bøgelund

AU - Grauslund, Morten

AU - Santoni-Rugiu, Eric

PY - 2015/11

Y1 - 2015/11

N2 - Background/Aim: We recently reported that miR-126 is down-regulated in malignant pleural mesothelioma (MPM) and can be combined into a 4-microRNA-classifier that can accurately diagnose MPM with high sensitivity and specificity. Herein we analyzed the epigenetic regulation of miR-126 and its host gene EGF-like domain, multiple 7 (EGFL7). Materials and Methods: Resected formalin-fixed paraffin-embedded MPM tissues from 29 patients, 14 patientmatched non-neoplastic pleura (NNP) specimens, 5 MPM diagnostic biopsies (DB), and 5 samples of pneumothoraxinduced benign reactive mesothelial proliferation (PTHX) were analyzed. miR-126 and EGFL7 mRNA were quantified by RT-qPCR. CpG-islands' methylation in the EGFL7 promoter was analyzed using methylation-specific PCR and in the MIR126-containing intron 7 was quantified by pyrosequencing. Results: Relative to NNP, EGFL7 was under-expressed more than 4-fold in MPM (p<0.001). EGFL7 mRNA and miR-126 levels correlated in MPM (p<0.01) and NNP (p<0.001). The EGFL7 promoter region was hypermethylated in 69% of MPM and 80% of DB samples, but not in NNP and PTHX samples. EGFL7 promoter hypermethylation was associated with epithelioid histology (p<0.05) and reduced patient-survival (p<0.05). Conclusion: In MPM, DNA-hypermethylation down-regulates miR-126 and its host gene EGFL7, therefore is a poor prognostic factor, and may represent a future therapeutic target for de-methylating strategies re-establishing EGFL7 and miR-126 expression.

AB - Background/Aim: We recently reported that miR-126 is down-regulated in malignant pleural mesothelioma (MPM) and can be combined into a 4-microRNA-classifier that can accurately diagnose MPM with high sensitivity and specificity. Herein we analyzed the epigenetic regulation of miR-126 and its host gene EGF-like domain, multiple 7 (EGFL7). Materials and Methods: Resected formalin-fixed paraffin-embedded MPM tissues from 29 patients, 14 patientmatched non-neoplastic pleura (NNP) specimens, 5 MPM diagnostic biopsies (DB), and 5 samples of pneumothoraxinduced benign reactive mesothelial proliferation (PTHX) were analyzed. miR-126 and EGFL7 mRNA were quantified by RT-qPCR. CpG-islands' methylation in the EGFL7 promoter was analyzed using methylation-specific PCR and in the MIR126-containing intron 7 was quantified by pyrosequencing. Results: Relative to NNP, EGFL7 was under-expressed more than 4-fold in MPM (p<0.001). EGFL7 mRNA and miR-126 levels correlated in MPM (p<0.01) and NNP (p<0.001). The EGFL7 promoter region was hypermethylated in 69% of MPM and 80% of DB samples, but not in NNP and PTHX samples. EGFL7 promoter hypermethylation was associated with epithelioid histology (p<0.05) and reduced patient-survival (p<0.05). Conclusion: In MPM, DNA-hypermethylation down-regulates miR-126 and its host gene EGFL7, therefore is a poor prognostic factor, and may represent a future therapeutic target for de-methylating strategies re-establishing EGFL7 and miR-126 expression.

KW - Adult

KW - Aged

KW - Biomarkers, Tumor

KW - Case-Control Studies

KW - DNA Methylation

KW - Endothelial Growth Factors

KW - Epigenesis, Genetic

KW - Female

KW - Follow-Up Studies

KW - Gene Expression Regulation, Neoplastic

KW - Humans

KW - Lung Neoplasms

KW - Male

KW - Mesothelioma

KW - MicroRNAs

KW - Middle Aged

KW - Neoplasm Staging

KW - Pleural Neoplasms

KW - Prognosis

KW - Promoter Regions, Genetic

KW - Real-Time Polymerase Chain Reaction

M3 - Journal article

C2 - 26504055

SN - 0250-7005

VL - 35

SP - 6223

EP - 6229

JO - Anticancer Research

JF - Anticancer Research

IS - 11

ER -

Methylation-associated Silencing of microRNA-126 and its Host Gene EGFL7 in Malignant Pleural Mesothelioma

Abstract

Keywords

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