Abstract
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.
Keywords: Dual-specificity phosphatase, miRNA, HaCaT, LPS, adalimumab, expression.
Graphical Abstract
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