S Garritano
A common polymorphism within MSLN Affects miR-611 binding site and soluble mesothelin levels in healthy people
Garritano, S; De Santi, C; Silvestri, R; Melaiu, O; Cipollini, M; Barone, E; Lucchi, M; Barale, R; Mutti, L; Gemignani, F; Bonotti, A; Foddis, R; Cristaudo, A; Landi, S
Authors
C De Santi
R Silvestri
O Melaiu
M Cipollini
E Barone
M Lucchi
R Barale
L Mutti
F Gemignani
A Bonotti
R Foddis
A Cristaudo
S Landi
Abstract
INTRODUCTION:
Soluble mesothelin related peptide (SMRP) was proposed as a promising diagnostic marker for malignant pleural mesothelioma (MPM). In a previous study, we found that rs1057147 within the 3' untranslated region of MSLN gene was associated with SMRP levels. Thus, we aimed to (1) confirm the previous association on a large series of volunteers and (2) test the hypothesis that the SNP could affect microRNA binding sites.
METHODS:
The association analysis was verified in 759 subjects. Then, in silico predictions highlighted miR-611 and miR-887 as candidate miRNAs binding to the polymorphic site. Thus, chimeric constructs bearing the alternative alleles (G > A) were assayed alone or in cotransfection with the miRNA mimics, with dual luciferase reporter assay in non-MPM Met-5A cells. The miRNAs were also assayed by western blot analysis for their ability to down-regulate endogenous mesothelin in the MPM Mero-14 cell line.
RESULTS:
We confirmed that, among non-MPM volunteers, GG homozygotes have the lowest SMRP levels. When the genotype is taken into account, the specificity of SMRP as biomarker improves from 79.7% to 85.3%. Dual-luciferase assays showed a significantly lower reporter activity when the vector harbored the G allele as compared to A allele. miR-887 mimic caused a reduced reporter activity of vectors harboring A or G alleles, while miR-611 was effective only on the vector harboring the G allele. Transfection of these miRNAs into Mero-14 cells significantly reduced endogenous MSLN protein.
CONCLUSION:
SMRP performance as diagnostic biomarker improved by considering the genotype rs1057147. This polymorphism most likely affects a binding site for miR-611.
Citation
Garritano, S., De Santi, C., Silvestri, R., Melaiu, O., Cipollini, M., Barone, E., …Landi, S. (2014). A common polymorphism within MSLN Affects miR-611 binding site and soluble mesothelin levels in healthy people. Journal of Thoracic Oncology, 9(11), 1662-1668. https://doi.org/10.1097/JTO.0000000000000322
Journal Article Type | Article |
---|---|
Online Publication Date | Nov 20, 2015 |
Publication Date | Nov 1, 2014 |
Deposit Date | Feb 2, 2015 |
Journal | Journal of Thoracic Oncology |
Print ISSN | 1556-0864 |
Electronic ISSN | 1556-1380 |
Publisher | Lippincott, Williams & Wilkins |
Peer Reviewed | Peer Reviewed |
Volume | 9 |
Issue | 11 |
Pages | 1662-1668 |
DOI | https://doi.org/10.1097/JTO.0000000000000322 |
Publisher URL | http://dx.doi.org/10.1097/JTO.0000000000000322 |
Related Public URLs | http://www.jto.org/ |
Additional Information | Funders : Buzzi Foundation (Italy) |
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