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首页> 外文期刊>Journal of experimental & clinical cancer research : >Exosomal miR-106b-5p derived from melanoma cell promotes primary melanocytes epithelial-mesenchymal transition through targeting EphA4
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Exosomal miR-106b-5p derived from melanoma cell promotes primary melanocytes epithelial-mesenchymal transition through targeting EphA4

机译：来自黑素瘤细胞的外泌体miR-106b-5p通过靶向epha4促进初级黑色细胞上皮 - 间充质转换

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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. 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. 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. Thus, melanoma cell-secreted exosomal miR-106b-5p may serve as a diagnostic indicator and potential therapeutic target in melanoma patients.
机译：癌症分泌的外泌体miRNA调节许多肿瘤的生物过程。黑色素瘤患者的外泌体miR-106b-5p的血清水平显着增加。然而，外泌体miR-106b-5p在黑素瘤中的作用和分子机制仍然不清楚。定量实时聚合酶链反应（QRT-PCR）用于检测黑素瘤组织中miR-106b-5p和epha4的表达。使用透射电子显微镜（TEM）和蛋白质印迹用于识别外鼻孔。 QRT-QPCR和Cy3标记的MiR-106B-5P用于证明黑素瘤细胞分泌的外泌体MIR-106B-5P的透射。用于蛋白质印迹，免疫荧光，粘附性，Transwell和划伤伤口测定用于探讨外泌体miR-106b-5p在黑素细胞中的作用。荧光素酶报告器测定和RNA-染色质免疫沉淀（芯片）测定用于确认产生促红细胞素的肝细胞癌受体A4（EphA4）是miR-106b-5p的直接靶标。我们发现，黑色素瘤组织中miR-106b-5p水平增加，高miR-106b-5p表达是黑素瘤患者整体存活的独立危险因素。 miR-106b-5p富集在黑色素瘤细胞分泌的外泌体中，并转移到黑色细胞中。外泌体miR-106b-5p促进了黑素细胞的上皮 - 间充质转换（EMT），迁移，侵袭和粘附性。 ExosoMal MiR-106B-5P通过靶向EPHA4来激活ERK途径的作用。我们证明外泌体MiR-106B-5P通过肺转移测定促进体内黑色素转移。因此，黑色素瘤细胞分泌的外泌体miR-106b-5p可以作为黑素瘤患者的诊断指示剂和潜在的治疗靶标。

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