Abstract
BackgroundCancer 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.MethodsInitially, 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.ResultsWe 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.ConclusionsOur results indicate that dysregulation of miRNA may play a role in the stem cell-like properties of ovarian CSCs.
Highlights
Cancer stem cells (CSCs) are thought to be a source of tumor recurrence due to their stem cell-like properties
Determination of the capacity of ovarian cancer cell lines for in vitro anchorage-independent growth We first tested the capacity for anchorage-independent growth, and tumor spheroid formation was noted in OVCAR3, TOV112D, and SKOV3 (Figure 1)
Spheroids are known to contain a greater number of CSCs [6], and in accordance with previous observations, increased paclitaxel resistance was noted in spherical ovarian cancer cells (Figure 2)
Summary
Cancer stem cells (CSCs) are thought to be a source of tumor recurrence due to their stem cell-like properties. The ability of cells to form tumor spheres is one such method, evaluating the capacity of cancer cells to grow as multi-cellular spheroids under non-differentiating and non-adherent conditions [5]. Using this method, ovarian CSCs from patients with ascites are first isolated and their ability to exhibit stem cell-like properties is examined [6]. In vivo serial transplantation assays, dye exclusion assays, and isolation via cell surface specific antigen profile methods are used to identify CSCs. In ovarian cancer, the most commonly used cell surface marker to identify ovarian CSCs involves the use of cluster of differentiation (CD) 133+ cell populations [7]
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