Changes in the Expression Pattern of DUSP1-7 and miRNA Regulating their Expression in the Keratinocytes Treated with
Beniamin Oskar Grabarek1,2,3, Maciej Dąbala4, Tomasz Kasela5
1Department of Histology, Cytophysiology, and Embryology in Zabrze, Faculty of Medicine in Zabrze, The University of Technology in Katowice, 41800, Zabrze, Poland.
Background:
Increased levels of phosphorylated ERK and p38 MAPK proteins have been observed in psoriatic skin biopsies compared to controls, which may be associated with an impaired expression pattern of dual activity protein phosphatase (DUSP).
Objective:
The purpose of this study was to assessment changes in expression profile of mRNA DUSP 1-7 and miRNA regulating their expression in human keratinocyte cells (HaCaT) had exposed to the liposaccharide A (LPS).
Methods:
HaCaT was exposed to 1 μg/ml LPS and next adalimumab by 2,8,24h compared to untreated cells. The microarray method was used to analyze expression pattern of mRNAs, miRNAs, and ELISA to evaluate changes in the level of the proteins. RTqPCR was used to validate the microarray data. Transcriptome Analysis Console and Statistica Software 13 PL were used in statistical analysis (p<0.05).
Results:
The highest changes in expression was observed for DUSP2 (FC +11.12) and DUSP5 (FC +5.53) in HaCaT culture after 2 hours exposition on adalimumab. It was observed that miR- 1275 (FC -2.39) and miR-34a (FC +6.52) might regulate level of DUSP2, and miR-27a (FC +3.55), miR-27b (FC +2.87) are involved in DUSP5 expression.
Conclusion:
The results obtained suggest that DUSP2 and DUSP5 may be considered as complementary molecular markers in the diagnosis and monitoring of the effectiveness of psoriasis therapy. It was confirmed that hsa-miR-34a, hsa-miR-1275, hsa-miR-3188, hsa-miR-382, hsa-miR- 27a, hsa-miR-27b, hsa-miR-16 have the highest influence on the expression pattern of DUSP1-7.
Insights
Dual-specificity phosphatases (DUSP) 2 and 5, and specific microRNAs, show altered expression in psoriatic keratinocytes. These findings suggest their potential as biomarkers for psoriasis diagnosis and therapy monitoring.
Area of Science:
- Dermatology
- Molecular Biology
- Biochemistry
Background:
- Psoriatic skin exhibits elevated phosphorylated ERK and p38 MAPK.
- This may correlate with altered expression of dual-specificity protein phosphatases (DUSP).
Purpose of the Study:
- To assess changes in mRNA expression of DUSP 1-7 and their regulating microRNAs.
- To investigate these changes in human keratinocyte cells (HaCaT) exposed to lipopolysaccharide A (LPS) and adalimumab.
Main Methods:
- HaCaT cells were treated with LPS and adalimumab.
- Microarray analysis was used for mRNA and miRNA expression profiling.
- ELISA and RTqPCR were employed for protein and data validation, respectively.
Main Results:
- Adalimumab treatment significantly altered DUSP2 (FC +11.12) and DUSP5 (FC +5.53) expression.
- Specific microRNAs, including miR-1275, miR-34a, miR-27a, and miR-27b, were identified as potential regulators of DUSP2 and DUSP5.
- The study identified key microRNAs influencing DUSP1-7 expression patterns.
Conclusions:
- DUSP2 and DUSP5 show promise as complementary biomarkers for psoriasis diagnosis and treatment monitoring.
- Several microRNAs (hsa-miR-34a, hsa-miR-1275, hsa-miR-27a, hsa-miR-27b, among others) significantly influence DUSP1-7 expression.
