Analysis of melanosomal miRNAs
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE72337 | GEO | 2016/08/09
SECONDARY ACCESSION(S): PRJNA293786
REPOSITORIES: GEO
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Project description:miRNA microarray profiling was performed for MNT-1 cells, pre-mature and mature melanosomes extracted from MNT-1 cells. In order to reveal the miRNAs that are most abundant in mature melanosomes, the most highly expressed miRNAs from mature melanosomes were compared with expression levels in pre-mature melanosomes. The analysis revealed five miRNAs that were highly expressed in mature melanosomes and had been previously associated with melanoma. Total RNA was extracted from MNT-1 cells (2 replicates), pre-mature and mature melanosomes. The melanosomes were either treated with RNase prior to RNA extraction (2 replicates each) or untreated (1 replicate each). Melanosomes were extracted from MNT-1 melanoma cells by sucrose-based density gradient ultracentrifugation. The treatment of melanosomes with RNase prior to RNA extraction was performed in order to remove any kind of RNA that may interact with or stick unspecifically to the outer side of the melanosome membrane. After 10 mins of RNAse treatment, RNase inhibitor was added. The samples of MNT-1 cells and untreated melanosomes were considered for the normalization, but not for further analyses. Microarray Unit of the Core Facility Genomics and Proteomics DKFZ
2016-08-09 | E-GEOD-72337 | biostudies-arrayexpress
Project description:miRNA microarray profiling was performed for exosomes and secreted melanosomes of MNT-1 cells as well as WM3682 and WM3314 cells and melanosomes isolated from cell homogenates. The aim of this study is to compare the miRNA content of exosomes and secreted melanosomes and to assess differences in the miRNA profiles of different melanoma cell lines and their melanosomes. Total RNA was extracted from MNT-1 exosomes and secreted melanosomes (2 replicates each) that were isolated from the conditioned medium of MNT-1 melanoma cells by differential ultracentrifugation. In addition, total RNA was isolated from WM3682 and WM3314 melanoma cells and melanosomes (1 replicate each). These latter melanosomes were isolated from cell homogenates by filtration through a 0.45 µm filter, followed by FACS sorting. Contributor: Microarray Unit of the Core Facility Genomics and Proteomics DKFZ
2016-08-09 | E-GEOD-79411 | biostudies-arrayexpress
Project description:mRNA profiling was performed on primary fibroblasts treated with melanosomes and untreated fibroblasts as control. The aim of this study was to explore whether melanoma-derived mature melanosomes functionally affect primary fibroblasts. A functional gene set enrichment analysis based on the resulting mRNA profiles predicted that an uptake of mature melanosomes by fibroblasts causes increased cell proliferation and cell motility. Expression profiling was performed for 3 replicates of primary human fibroblasts that were treated with mature melanosomes, 1 replicate of fibroblasts treated with early melanosomes and 3 replicates of untreated fibroblasts as controls. Melanosomes were extracted from the human melanoma cell line MNT-1. Fibroblasts were treated with melanosomes for 24h. Fold changes were calculed for fibroblasts treated with mature melanosomes vs. controls. The sample of fibroblasts treated with early melanosomes was considered for the normalization, but not for further analyses. Microarray Unit of the Core Facility Genomics and Proteomics DKFZ
2016-08-09 | E-GEOD-72229 | biostudies-arrayexpress
Project description:miRNA microarray profiling was performed for exosomes and secreted melanosomes of MNT-1 cells as well as WM3682 and WM3314 cells and melanosomes isolated from cell homogenates. The aim of this study is to compare the miRNA content of exosomes and secreted melanosomes and to assess differences in the miRNA profiles of different melanoma cell lines and their melanosomes.
2016-08-09 | GSE79411 | GEO
Project description:mRNA profiling was performed on primary fibroblasts treated with melanosomes and untreated fibroblasts as control. The aim of this study was to explore whether melanoma-derived mature melanosomes functionally affect primary fibroblasts. A functional gene set enrichment analysis based on the resulting mRNA profiles predicted that an uptake of mature melanosomes by fibroblasts causes increased cell proliferation and cell motility.
2016-08-09 | GSE72229 | GEO
Project description:mRNA profiles generated from primary fibroblast upon treatment with miR-211, miR-302 or melanoma melanosomes. Abstract: Melanoma originates in the epidermis and enters the metastatic and lethal phase upon invasion into the dermis. However, the interactions between melanoma cells and the dermis prior to this invasion have been poorly studied. Here we uncover that melanoma cells directly affect the formation of the dermal tumor niche by microRNA (miRNA) trafficking prior to invading the dermis. Melanocytes, the cells of melanoma origin, are specialized in trafficking of pigment vesicles, termed melanosomes and, interestingly, melanoma cells retain this trafficking ability. In melanoma in-situ specimens, we found melanosome markers in distal fibroblasts prior to the invasion of melanoma cells into the dermis. Melanoma-derived melanosomes carry miRNAs into primary fibroblasts that trigger changes in the fibroblasts, including increased proliferation, migration, and expression of pro-inflammatory genes, all known features of cancer-associated fibroblasts (CAFs). Specifically, we found that melanosomal miRNA-211 directly targets IGF2R and leads to MAPK signaling activation in fibroblasts, which reciprocally encourages melanoma growth. Treatment of melanoma cells with a melanosome release-inhibitor prevented CAF formation. Since the first interaction of melanoma cells with blood vessels occurs in the dermis, our data suggest a promising opportunity to block melanoma cell invasion by preventing the formation of the dermal tumor niche. In the paper we showed the 10% of most differentially expressed mRNA upon miR-211, miR-320c and melanosomes treatment and overlap of 2000 downregulated mRNA upon miR-211 and melanosomes treatment with predicted target gene miR-211 and CAFs related genes Expresssion profiling was performed for primary fibroblasts transfected with miRNA-211 mimic and miRNA-320c mimic.
2016-08-09 | E-GEOD-72619 | biostudies-arrayexpress
Project description:Melanoma-derived melanosomes traffic mature let-7i to lymphatic endothelial cells, which mediate pro-lymphangiogenic phenotypic changes by the induction of type I interferon signaling. Further, transcriptome analysis upon treatments with melanosomes or let-7i reveal the enhancement of IFI6 expression in lymphatic cells.
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Project description:mRNA profiles generated from primary fibroblast upon treatment with miR-211, miR-302 or melanoma melanosomes. Abstract: Melanoma originates in the epidermis and enters the metastatic and lethal phase upon invasion into the dermis. However, the interactions between melanoma cells and the dermis prior to this invasion have been poorly studied. Here we uncover that melanoma cells directly affect the formation of the dermal tumor niche by microRNA (miRNA) trafficking prior to invading the dermis. Melanocytes, the cells of melanoma origin, are specialized in trafficking of pigment vesicles, termed melanosomes and, interestingly, melanoma cells retain this trafficking ability. In melanoma in-situ specimens, we found melanosome markers in distal fibroblasts prior to the invasion of melanoma cells into the dermis. Melanoma-derived melanosomes carry miRNAs into primary fibroblasts that trigger changes in the fibroblasts, including increased proliferation, migration, and expression of pro-inflammatory genes, all known features of cancer-associated fibroblasts (CAFs). Specifically, we found that melanosomal miRNA-211 directly targets IGF2R and leads to MAPK signaling activation in fibroblasts, which reciprocally encourages melanoma growth. Treatment of melanoma cells with a melanosome release-inhibitor prevented CAF formation. Since the first interaction of melanoma cells with blood vessels occurs in the dermis, our data suggest a promising opportunity to block melanoma cell invasion by preventing the formation of the dermal tumor niche. In the paper we showed the 10% of most differentially expressed mRNA upon miR-211, miR-320c and melanosomes treatment and overlap of 2000 downregulated mRNA upon miR-211 and melanosomes treatment with predicted target gene miR-211 and CAFs related genes
2016-08-09 | GSE72619 | GEO
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