Related Experiment Video
Updated: May 19, 2026
Measuring Retinal Vessel Diameter from Mouse Fluorescent Angiography Images
Published on: May 19, 2023
The changes of microRNA expression profiles and tyrosinase related proteins in MITF knocked down melanocytes
Ping Wang1, Yong Li, Weisong Hong
1Department of Dermatology, the Affiliated 3rd Hospital of Hangzhou, Anhui Medical University, Hangzhou, 310009, China. dermwang@yahoo.com.cn
Abstract:
Microphthalmia-associated transcription factor (MITF) is a master regulator in melanocyte proliferation, development, survival and melanoma formation. In melanocyte dysfunction disease, it is observed that the expressions of MITF, tyrosinase (TYR), tyrosinase related protein 1 (TYRP1) and tyrosinase related protein 2 (TYRP2)/dopachrome tautomerase (DCT) are changed, the consequence of which remains unclear. In this study, we focused on the change of microRNA (miRNA) profiles and Tyrosinase Related Proteins (TRPs) in MITF knocked down melanocytes. For the first time, we assayed the MITF-KD miRNA profiles using a miRNA microarray and found that hsa-miR-1225-3p, hsa-miR-634, hsa-miR-197, hsa-miR-766, hsa-miR-574-5p and hsa-miR-328 were upregulated, and hsa-miR-720 and hsa-miR-1308 were downregulated in MITF knocked down melanocytes. These miRNAs were validated by miRNA real time qPCR. These miRNA potential targets, especially the TRPs, were analyzed according to the miRNA database (Sanger Center). By TargetScan prediction, the hsa-miR-634 and hsa-miR-328 have poorly conserved sites on TYR and hsa-miR-197 have poorly conserved sites on TYR1. Through qPCR and western blotting we found that the expression of TYR and TYRP1 were dramatically decreased and the expression of TYRP2 was increased in MITF knocked down melanocytes (MITF-KD). These results suggested that the miRNAs may be involved in MITF regulation of TYR, TYRP1 and TYRP2, which provides a new clue for understanding the role of miRNAs in melanocyte dysfunctional disease.
Insights
Microphthalmia-associated transcription factor (MITF) knockdown alters microRNA profiles in melanocytes. This impacts Tyrosinase Related Proteins (TRPs), offering insights into melanocyte dysfunction diseases.
Area of Science:
- Cell Biology
- Molecular Biology
- Genetics
Background:
- Microphthalmia-associated transcription factor (MITF) is crucial for melanocyte function and melanoma.
- Dysregulation of MITF and Tyrosinase Related Proteins (TRPs) is observed in melanocyte disorders.
- The role of microRNAs (miRNAs) in MITF-mediated regulation of TRPs is not fully understood.
Purpose of the Study:
- To investigate the impact of MITF knockdown on miRNA expression profiles in melanocytes.
- To identify specific miRNAs affected by MITF knockdown.
- To analyze the relationship between altered miRNAs and the expression of Tyrosinase Related Proteins (TYR, TYRP1, TYRP2).
Main Methods:
- Melanocyte cell culture with MITF knockdown (MITF-KD).
- miRNA microarray analysis to profile miRNA expression.
- Quantitative real-time PCR (qPCR) for miRNA validation.
- Bioinformatic analysis (Sanger Center, TargetScan) to predict miRNA targets.
- qPCR and Western blotting to assess TRP expression levels.
Main Results:
- MITF knockdown led to significant changes in miRNA expression, with several miRNAs upregulated (e.g., hsa-miR-1225-3p, hsa-miR-634) and others downregulated (e.g., hsa-miR-720).
- Bioinformatic predictions suggested potential interactions between specific miRNAs (hsa-miR-634, hsa-miR-328, hsa-miR-197) and TRP genes.
- Experimental validation showed decreased expression of TYR and TYRP1, and increased expression of TYRP2 in MITF-KD melanocytes.
Conclusions:
- MITF knockdown significantly alters miRNA expression in melanocytes.
- These altered miRNAs are potentially involved in regulating the expression of TYR, TYRP1, and TYRP2.
- The findings provide new insights into the role of miRNAs in MITF-mediated melanocyte regulation and dysfunction.
Related Concept Videos
01:22MicroRNAs
01:22MicroRNAs