MicroRNA-142-5p contributes to Hashimoto’s thyroiditis by targeting CLDN1Report as inadecuate




MicroRNA-142-5p contributes to Hashimoto’s thyroiditis by targeting CLDN1 - Download this document for free, or read online. Document in PDF available to download.

Journal of Translational Medicine

, 14:166

DiseaseBiomarkers

Abstract

BackgroundMicroRNAs have the potential as diagnostic biomarkers and therapeutic targets in autoimmune diseases. However, very limited studies have evaluated the expression of microRNA profile in thyroid gland related to Hashimoto’s thyroiditis HT.

MethodsMicroRNA microarray expression profiling was performed and validated by quantitative RT-PCR. The expression pattern of miR-142-5p was detected using locked nucleic acid-in situ hybridization. The target gene was predicted and validated using miRNA targets prediction database, gene expression analysis, quantitative RT-PCR, western blot, and luciferase assay. The potential mechanisms of miR-142-5p were studied using immunohistochemistry, immunofluorescence, and quantitative assay of thyrocyte permeability.

ResultsThirty-nine microRNAs were differentially expressed in HT Fold change ≥2, P < 0.05 and miR-142-5p, miR-142-3p, and miR-146a were only high expression in HT thyroid gland P < 0.001. miR-142-5p, which was expressed at high levels in injured follicular epithelial cells, was also detected in HT patient serum and positively correlated with thyroglobulin antibody r ≥ 0.6, P < 0.05. Furthermore, luciferase assay demonstrated CLDN1 was the direct target gene of miR-142-5p P < 0.05, and Immunohistochemical staining showed a reverse expression patterns with miR-142-5p and CLDN1. Overexpression of miR-142-5p in thyrocytes resulted in reducing of the expression of claudin-1 both in mRNA and protein level P = 0.032 and P = 0.009 respectively and increasing the permeability of thyrocytes monolayer P < 0.01.

ConclusionsOur findings indicate a previously unrecognized mechanism that miR-142-5p, targeting CLDN1, plays an important role in HT pathogenesis.

KeywordsAutoimmune diseases Hashimoto’s thyroiditis miRNA miR-142-5p CLDN1 AbbreviationsRT-PCRreal time-polymerase chain reaction

CLDN1claudin-1

HTHashimoto’s thyroiditis

UTRsuntranslated regions

FFPEformalin-fixed paraffin-embedded

PTCpapillary thyroid carcinoma

LNAlocked nucleic acid

ISHin situ hybridization

PBSphosphate buffered saline

FITCfluorescein isothiocyanate

BSAbovine serum albumin

SSCsaline sodium citrate

TBSTtris buffered saline with tween

DABdiaminobenzidine

TgAbthyroglobulin antibody

TPOAbperoxidase antibody

TSHthyroid-stimulating hormone

Tgthyroglobulin

LVlentivirus vector

EGFPenhanced green fluorescent protein

RFUrelative fluorescence unit

Jin Zhu and Yuehua Zhang contributed equally to this work

Electronic supplementary materialThe online version of this article doi:10.1186-s12967-016-0917-6 contains supplementary material, which is available to authorized users.

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