Wenkang Luan1, Yuting Ding2, Haolan Xi3, Hongru Ruan4, Feng Lu4, Shaojun Ma4, Jinlong Wang4. 1. Department of Plastic Surgery, Affiliated People's Hospital of Jiangsu University, 8 Dianli Road, Zhenjiang, 212000, Jiangsu, China. luanwenkang@126.com. 2. Department of Rehabilitation, Changshu No. 2 People's Hospital (The 5th Clinical Medical College of Yangzhou University), Changshu, Jiangsu, China. 3. Department of Ophthalmology, Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu, China. 4. Department of Plastic Surgery, Affiliated People's Hospital of Jiangsu University, 8 Dianli Road, Zhenjiang, 212000, Jiangsu, China.
Abstract
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.
BACKGROUND:Cancer-secreted exosomal miRNAs regulates the biological processes of many tumours. The serum level of exosomal miR-106b-5p is significantly increased in melanomapatients. 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 melanomapatients.
Authors: D Michiel Pegtel; Katherine Cosmopoulos; David A Thorley-Lawson; Monique A J van Eijndhoven; Erik S Hopmans; Jelle L Lindenberg; Tanja D de Gruijl; Thomas Würdinger; Jaap M Middeldorp Journal: Proc Natl Acad Sci U S A Date: 2010-03-18 Impact factor: 11.205