miR-331-5p Affects Motility of Thyroid Cancer Cell Lines and Regulates BID Expression

Author:

Orlandella Francesca Maria12,Imperlini Esther3ORCID,Pane Katia4ORCID,Luciano Neila5ORCID,Braile Mariantonia4,De Stefano Anna Elisa12,Iervolino Paola Lucia Chiara5,Ruocco Alessandro26ORCID,Orrù Stefania12,Franzese Monica4ORCID,Salvatore Giuliana12

Affiliation:

1. Dipartimento delle Scienze Mediche, Motorie e del Benessere, Università degli Studi di Napoli “Parthenope”, 80133 Naples, Italy

2. CEINGE Biotecnologie Avanzate Franco Salvatore S.c.a.r.l, 80145 Naples, Italy

3. Department for Innovation in Biological, Agrofood and Forest Systems, University of Tuscia, 01100 Viterbo, Italy

4. IRCCS SYNLAB SDN, 80143 Naples, Italy

5. Dipartimento di Scienze Biomediche Avanzate, Università degli Studi di Napoli “Federico II”, 80131 Naples, Italy

6. Dipartimento di Biologia, Università degli Studi di Napoli “Federico II”, 80126 Naples, Italy

Abstract

During tumorigenesis, miRNAs with unbalanced expression profiles can increase the threat of disease progression. Here, we focus on the role of miR-331-5p in the pathogenesis of thyroid cancer (TC). In vitro studies were conducted using TC cell lines after the forced expression and silencing of miR-331-5p. Cell proliferation and viability were analyzed via cell counts and colorimetric assays. Cell motility was analyzed via wound healing assays, Transwell migration and invasion assays, and Matrigel Matrix assays. The putative targets of miR-331-5p were unveiled via label-free proteomic screening and then verified using Western blot and luciferase assays. Expression studies were conducted by interrogating The Cancer Genome Atlas (TCGA). We found that ectopic miR-331-5p expression reduces TC cell motility, while miR-331-5p silencing induces the opposite phenotype. Proteomic screening revealed eight putative downregulated targets of miR-331-5p, among which BID was confirmed as a direct target. TCGA data showed the downregulation of miR-331-5p and the upregulation of BID in TC tissues. In summary, deregulation of the miR-331-5p/BID axis could enhance the aggressiveness of TC cell lines, providing new insights into the mechanisms of the progression of this disease and suggesting a potential role of the component factors as possible biomarkers in TC tissues.

Funder

Ministero dell’Istruzione, dell’Università e della Ricerca bando PRIN

MUR, PNR

Publisher

MDPI AG

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