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

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Authors: Eun Ji Nam ; Maria Lee ; Ga Won Yim ; Jae Hoon Kim ; Sunghoon Kim ; Sang Wun Kim ; Young Tae Kim

MeSH: AC133 Antigen ; Antigens, CD/metabolism* ; Blotting, Western ; Cell Proliferation/drug effects ; Cluster Analysis ; Drug Resistance, Neoplasm/drug effects ; Drug Resistance, Neoplasm/genetics ; Female ; Flow Cytometry ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Glycoproteins/metabolism* ; Humans ; MicroRNAs/genetics* ; MicroRNAs/metabolism ; Neoplastic Stem Cells/drug effects ; Neoplastic Stem Cells/metabolism ; Neoplastic Stem Cells/pathology ; Oligonucleotide Array Sequence Analysis ; Ovarian Neoplasms/genetics* ; Ovarian Neoplasms/pathology* ; Paclitaxel/pharmacology ; Peptides/metabolism* ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; Spheroids, Cellular/drug effects ; Spheroids, Cellular/metabolism* ; Spheroids, Cellular/pathology* ; Tumor Cells, Cultured

Keywords: MicroRNA ; Cancer stem cell ; Ovarian cancer ; CD133 ; OVCAR3 ; Chemoresistance

Abstract: BACKGROUND: 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.

METHODS: 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.

RESULTS: 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.

CONCLUSIONS: Our results indicate that dysregulation of miRNA may play a role in the stem cell-like properties of ovarian CSCs.