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MicroRNA profiling of a CD133(+) spheroid-forming subpopulation of the OVCAR3 human ovarian cancer cell line.

ABSTRACT: Cancer stem cells (CSCs) are thought to be a source of tumor recurrence due to their stem cell-like properties. MicroRNAs (miRNAs) regulate both normal stem cells and CSCs, and dysregulation of miRNAs has an important role in tumorigenesis. Cluster of differentiation (CD) 133(+) and spheroid formation have been reported to be one of the main features of ovarian CSCs. Therefore, we determined the miRNA expression profile of a CD133(+) spheroid-forming subpopulation of the OVCAR3 human ovarian cancer cell line.Initially, we confirmed the enrichment of the OVCAR3 CD133 subpopulation by evaluating in vitro anchorage-independent growth. After obtaining a subpopulation of CD133(+) OVCAR3 cells with?>?98% purity via cell sorting, miRNA microarray and real-time reverse transcription-polymerase chain reaction (RT-PCR) were performed to evaluate its miRNA profile.We found 37 differentially expressed miRNAs in the CD133(+) spheroid-forming subpopulation of OVCAR3 cells, 34 of which were significantly up-regulated, including miR-205, miR-146a, miR-200a, miR-200b, and miR-3, and 3 of which were significantly down-regulated, including miR-1202 and miR-1181.Our results indicate that dysregulation of miRNA may play a role in the stem cell-like properties of ovarian CSCs.

SUBMITTER: Nam EJ

PROVIDER: S-EPMC3480901 | biostudies-other | 2012 May

REPOSITORIES: biostudies-other

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MicroRNA profiling of a CD133(+) spheroid-forming subpopulation of the OVCAR3 human ovarian cancer cell line.

Nam Eun Ji EJ Lee Maria M Yim Ga Won GW Kim Jae Hoon JH Kim Sunghoon S Kim Sang Wun SW Kim Young Tae YT

BMC medical genomics 20120529

<h4>Background</h4>Cancer stem cells (CSCs) are thought to be a source of tumor recurrence due to their stem cell-like properties. MicroRNAs (miRNAs) regulate both normal stem cells and CSCs, and dysregulation of miRNAs has an important role in tumorigenesis. Cluster of differentiation (CD) 133(+) and spheroid formation have been reported to be one of the main features of ovarian CSCs. Therefore, we determined the miRNA expression profile of a CD133(+) spheroid-forming subpopulation of the OVCAR ...[more]

PMID: 22643117

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Project description:Osteosarcoma (OS) is a primary malignant tumor of bone occurring in young adults. OS stem cells (OSCs) play an important role in the occurrence, growth, metastasis, drug resistance and recurrence of OS. CD133 is an integral membrane glycoprotein, which has been identified as an OSC marker. However, the mechanisms of metastasis, chemoresistance, and progression in CD133(+) OSCs need to be further explored. In this study, we aim to explore differences in miRNA levels between CD133(+) and CD133(-) cells from the MG-63 cell line. We found 20 differentially expressed miRNAs (DEmiRNAs) (16 upregulated and 4 downregulated) in CD133(+) cells compared with CD133(-) cells. Hsa-miR-4485-3p, hsa-miR-4284 and hsa-miR-3656 were the top three upregulated DEmiRNAs, while hsa-miR-487b-3p, hsa-miR-493-5p and hsa-miR-431-5p were the top three downregulated DEmiRNAs. In addition, RT-PCR analysis confirmed that the expression levels of hsa-miR-4284, hsa-miR-4485-3p and hsa-miR-3656 were significantly increased, while the expression levels of hsa-miR-487b-3p, hsa-miR-493-5p, and hsa-miR-431-5p were significantly decreased in CD133(+) cells compared with CD133(-) cells. Moreover, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that predicted or validated target genes for all 20 DEmiRNAs or the selected 6 DEmiRNAs participated in the "PI3K-Akt signaling pathway," "Wnt signaling pathway," "Rap1 signaling pathway," "Cell cycle" and "MAPK signaling pathway". Among the selected six DEmiRNAs, miR-4284 was especially interesting. MiR-4284 knockdown significantly reduced the sphere forming capacity of CD133(+) OS cells. The number of invasive CD133(+) OS cells was markedly decreased after miR-4284 knockdown. In addition, miR-4284 knockdown increased the p-β-catenin levels in CD133(+) OS cells. In conclusion, RNA-seq analysis revealed DEmiRNAs between CD133(+) and CD133(-) cells. MiRNAs might play significant roles in the function of OSCs and could serve as targets for OS treatment. MiR-4284 prompted the self-renewal and invasion of OSCs. The function of miR-4284 might be associated with the Wnt signaling pathway.

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Dataset Information

MicroRNA profiling of a CD133(+) spheroid-forming subpopulation of the OVCAR3 human ovarian cancer cell line.

Publications

MicroRNA profiling of a CD133(+) spheroid-forming subpopulation of the OVCAR3 human ovarian cancer cell line.

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