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Published on: June 7, 2019
Exosomal miR-106b-5p derived from melanoma cell promotes primary melanocytes epithelial-mesenchymal transition
Wenkang Luan1, Yuting Ding2, Haolan Xi3
1Department of Plastic Surgery, Affiliated People's Hospital of Jiangsu University, 8 Dianli Road, Zhenjiang, 212000, Jiangsu, China. luanwenkang@126.com.
Background:
Cancer-secreted exosomal miRNAs regulates the biological processes of many tumours. The serum level of exosomal miR-106b-5p is significantly increased in melanoma patients. However, the role and molecular mechanisms of exosomal miR-106b-5p in melanoma remains unclear.
Methods:
Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-106b-5p and EphA4 in melanoma tissues. Transmission electron microscopy (TEM) and western blotting were used to identify exosome. QRT-qPCR and Cy3-labelled miR-106b-5p were used to demonstrated the transmission of melanoma cell-secreted exosomal miR-106b-5p. Western blotting, Immunofluorescence, adhesion, transwell and scratch wound assay were used to explore the role of exosomal miR-106b-5p in melanocytes. Luciferase reporter assays and RNA-Chromatin Immunoprecipitation (ChIP) assay were used to confirm whether erythropoietin-producing hepatocellular carcinoma receptor A4 (EphA4) was a direct target of miR-106b-5p.
Results:
We found that miR-106b-5p levels were increased in melanoma tissue, and high miR-106b-5p expression is an independent risk factor for the overall survival of patients with melanoma. miR-106b-5p is enriched in melanoma cell-secreted exosomes and transferred to melanocytes. Exosomal miR-106b-5p promotes the epithelial-to-mesenchymal transition (EMT), migration, invasion and adhesion of melanocytes. Exosomal miR-106b-5p exerted its role by targeting EphA4 to activate the ERK pathway. We demonstrated that exosomal miR-106b-5p promoted melanoma metastasis in vivo through pulmonary metastasis assay.
Conclusions:
Thus, melanoma cell-secreted exosomal miR-106b-5p may serve as a diagnostic indicator and potential therapeutic target in melanoma patients.
Insights
Melanoma patients have increased exosomal miR-106b-5p, which promotes cancer cell migration and invasion. This microRNA may serve as a diagnostic marker and therapeutic target for melanoma.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Exosomal microRNAs (miRNAs) secreted by cancer cells influence tumor progression.
- Serum levels of exosomal miR-106b-5p are elevated in melanoma patients.
- The specific role and mechanisms of exosomal miR-106b-5p in melanoma remain largely unknown.
Purpose of the Study:
- To investigate the role and molecular mechanisms of exosomal miR-106b-5p in melanoma.
- To determine if exosomal miR-106b-5p can be a diagnostic indicator for melanoma.
Main Methods:
- Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were used to analyze miR-106b-5p and EphA4 expression.
- Exosomes were identified using transmission electron microscopy (TEM).
- Functional assays (adhesion, transwell, scratch wound) and molecular analyses (luciferase reporter, ChIP) were performed to elucidate the mechanism of action and target validation.
Main Results:
- Increased miR-106b-5p levels in melanoma tissues correlated with poorer patient survival.
- Melanoma cell-secreted exosomes enriched with miR-106b-5p were transferred to melanocytes.
- Exosomal miR-106b-5p promoted melanocyte epithelial-to-mesenchymal transition (EMT), migration, invasion, and adhesion by targeting EphA4 and activating the ERK pathway.
- In vivo studies confirmed that exosomal miR-106b-5p promotes melanoma metastasis.
Conclusions:
- Melanoma cell-secreted exosomal miR-106b-5p plays a crucial role in promoting melanoma progression and metastasis.
- Exosomal miR-106b-5p shows potential as a diagnostic biomarker for melanoma.
- Targeting exosomal miR-106b-5p represents a promising therapeutic strategy for melanoma treatment.
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