Target regulation of micro-RNA-203 to the expression of collagen type Ⅳ alpha 4 and its role in oral submucous fibrosis
10.3760/cma.j.issn.1002-0098.2016.09.004
- VernacularTitle:微小RNA-203对Ⅳ型胶原蛋白α4链基因的靶向调控及其在口腔黏膜下纤维化中的作用
- Author:
Jun CHEN
1
;
Binjie LIU
;
Cui DU
;
Qiong CAO
;
Min LI
;
Hui FENG
Author Information
1. 中南大学湘雅口腔医院黏膜科
- Keywords:
Oral submucous fibrosis;
MicroRNA;
Genes,reporter;
Collagen type Ⅳ alpha 4
- From:
Chinese Journal of Stomatology
2016;51(9):526-531
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of microRNA-203(miR-203) on the expression of collagen type Ⅳ alpha 4(COL4A4) and its role in oral submucous fibrosis(OSF).Methods It has been revealed that COL4A4 is the putative target of miR-203 by retrieving a variety of bioinformatics software tools.The expression of COL4A4 protein in the indicated tissues was examined by Western blotting analysis and the potential binding sites of miR-203 and COL4A4-3'UTR was analyzed by using bioinformatics.This study integrated a fragment of COL4A4 3'UTR containing the target sequence into the pMIR-REPORT luciferase vector.Then,the COL4A4 3'UTR-luciferase and miR-203 were contransfected into 293T cells.Beta-gal was used as a control to monitor transfection efficiency.Cells were harvested and luciferase activity was measured after 24 h of transfection.Results The relative expression mean valves of COL4A4 were 2.46±2.26 in OSF and 0.70±0.49 in normal control.The expression of COL4A4 in OSF was higher than normal control(P＜0.05) and the difference between groups was statistically significant.The relative expression mean valves of miR-203 were 2.57±2.09 in OSF and 154.07±191.11 in normal control.The expression of miR-203 in OSF was lower than in normal control(P＜0.01) and the difference between groups was statistically significant.Bioinformatics analysis revealed that the COL4A4 3'UTR had a conserved site (site 1) and 3 non-conserved sites(site 2,site 3,site4) complementary to the miR-203.MiR-203 could significantly inhibit site 1-luciferase reporter activity,partially inhibited site 3-and site 4-1uciferase reporter activities,while no significant effect on site 3-luciferase reporter activity.Conclusions MiR-203 was negatively correlated with the expression of COL4A4 protein in OSF.Moreover,miR-203 repressed the expression of COL4A4 by targeting 3'UTR site of COL4A4 mRNA.
