Downregulation of miR29b3p promotes αtubulin deacetylation by targeting the interaction of matrix metalloproteinase9 with integrin ß1 in nasal polyps.
Int J Mol Med
; 48(1)2021 07.
Article
em En
| MEDLINE
| ID: mdl-33982786
ABSTRACT
Matrix metalloproteinase (MMP)9 is a key enzyme responsible for extracellular matrix degradation and contributes to the progressive histological changes observed in lower respiratory tract infections. Integrin ß1 and αtubulin are potential MMP9interacting proteins, and microRNA (miR)29b3p can regulate MMP9 expression. MMP9 is highly expressed in chronic rhinosinusitis with nasal polyps (CRSwNPs), regardless of its effects on miR29b3p, integrin ß1 and αtubulin expression. In the present study, samples from 100 patients with CRSwNPs were examined via reverse transcriptionquantitative PCR to assess the mRNA expression of miR29b3p, and western blotting was performed to assess the protein expression of MMP2, MMP9, acetylαtubulin, integrin ß1 and tissue inhibitor of metalloproteinase 1 (TIMP1). A dualluciferase reporter assay was used to verify the direct binding of miR29b3p and MMP2/MMP9. Coimmunoprecipitation (CoIP) and GST pulldown assays showed that integrin ß1 and αtubulin were MMP9interacting proteins. Cell viability, apoptosis and inflammatory cytokine levels were determined via a Cell Counting Kit8 assay, flow cytometry and ELISA, respectively. miR29b3p expression was found to be positively correlated with MMP2 and MMP9 expression. Whereas, TIMP1 expression was negatively correlated with MMP2 and MMP9 expression. The dualluciferase assay revealed that miR29b3p targeted the 3' untranslated region of MMP2/MMP9. The CoIP and GST pulldown assays showed that MMP9 could directly bind to integrin ß1 and indirectly bind to αtubulin. Finally, the overexpression of miR29b3p decreased the expression of MMP9 and increased the levels of acetylαtubulin. By contrast, the knockdown of miR29b3p increased the expression of MMP9 and decreased the levels of acetylαtubulin. Additionally, MMP9 expression was found to be negatively correlated with acetylαtubulin expression. Of note, the expression of integrin ß1 did not change following the overexpression and knockdown of MMP9. Finally, the overexpression of miR29b3p not only decreased MMP9 expression, but also alleviated lipopolysaccharideinduced inflammation in NP69 cells. The results showed that the downregulation of miR29b3p promoted αtubulin deacetylation by increasing the number of MMP9integrin ß1 complexes in CRSwNPs, thus targeting miR29b3p/MMP9 may be a potential novel strategy for the clinical treatment of CRSwNPs.
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Base de dados:
MEDLINE
Assunto principal:
Sinusite
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Tubulina (Proteína)
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Pólipos Nasais
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Integrina beta1
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Metaloproteinase 9 da Matriz
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article