Regulation of miR140-5p for paraoxonase 1 expression in HepG2 cells and its clinical application
10.13602/j.cnki.jcls.2019.02.14
- VernacularTitle:MiR140-5p在HepG2细胞中对对氧磷酶1表达的调控及其临床应用
- Author:
Jiaxing LIU
1
;
Bing WANG
2
;
Jicheng XING
2
;
Yujie HE
2
;
Aizhen YANG
3
;
Hong QIU
2
Author Information
1. Bayi College of Clinical Medicine, Anhui Medical University
2. Department of Clinical Laboratory, the 81st Hospital of PLA
3. Department of central Laboratory, the 81st Hospital of PLA
- Publication Type:Journal Article
- Keywords:
miR140-5p;
paraoxonase 1;
HepG2 cell;
nonalcoholic steatohepatitis
- From:
Chinese Journal of Clinical Laboratory Science
2019;37(2):137-141
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of miRNA on the expression of paraoxonase 1 (PON1) and its clinical application in the patients with nonalcoholic steatohepatitis (NASH).
Methods:Bioinformatics methods were used to analyze and predict PON1 related regulation on miRNA. PON1 luciferase reporter gene vectors were constructed and the activity of dual luciferase was analyzed. The up/down-regulated levels of miRNA in HepG2 cells of different groups were detected by real-time fluorescence quantitative PCR (qRT-PCR), and the levels of PON1 protein in HepG2 cells were detected by western blot. The levels of miR140-5p in the serum of healthy people and NASH patients were also analyzed by qRT-PCR.
Results:According to the prediction of TargetScan database, miR140-5p may bind complementarily to the end of PON13′-UTR. The analysis for the activity of dual luciferase reporter gene showed that miR-140-5p mimic significantly downregulated the fluorescence of wild type PON1 vector (P＜0.01). The results of qRT-PCR demonstrated that miR-140-5p mimic group showed high overexpression (P＜0.01) compared with the normal cell control group and the negative mimic control group, while miR-140-5p inhibitor group appeared corresponding low expression (P＜0.05). western blot results suggested that the transfection of miR140-5p mimic significantly down-regulated the expression of PON1 (P＜0.01) while miR140-5p inhibitor up-regulated this expression (P＜0.01). Compared with the healthy control group, the level of miR140-5p was decreased in the serum of NASH patients, and the difference was statistically significant (P＜0.01).
Conclusion:miR140-5p may be involved in the progression of nonalcoholic steatohepatitis through regulation for the posttranscriptional gene expression of PON1.
