Department of Obstetrics and Gynecology, The Third Affiliated Hospital, Guangzhou Medical University, Guangzhou, P.R. China.
Oncol Rep. 2011 Jan;25(1):141-6.
Cancer stem cells (CSCs) form a very rare population within tumors and possess the ability to proliferate and self-renew indefinitely. The cluster of differentiation (CD) 133+ ovarian CSCs (OCSCs) have been identified recently and their clinical implications are about to be clarified. In this context, we use the CD133 antigen as a marker of OCSCs in OVCAR3 cells and show that microRNAs (miRNAs) are aberrantly expressed in this subpopulation. The OCSCs in the OVCAR3 cell line were identified by the monoclonal mouse anti-CD133-1 antibody (clone CD133). Microarray and real-time reverse transcription-polymerase chain reaction (RT-PCR) analyses were used to identify miRNAs with altered expression in CD133+ cells. The expression levels of dysregulated miRNAs, namely, miR-204, miR-206, miR-223, miR-9, miR-100, and miR-200c, were examined using TaqMan PCR. The RNA and protein levels of stem cell-specific genes were examined by real-time RT-PCR and western blot analyses. Our results of microarray and real-time RT-PCR analyses revealed distinct miRNA expression profiles between CD133+ and CD133- OVCAR3 cells. The expression of stem cell-specific genes, namely, Oct3/4, Sox2, and Nanog, was higher in CD133+ cells than in CD133- cells while that of FoxD3, was lower in CD133+ cells than in CD133- cells. In conclusion, our data indicate that CD133 expression defines a tumor-initiating subpopulation of cells in the OVCAR3 cell line. The overall miRNA expression profile of CD133+ OVCAR3 cells was clearly distinct from that of CD133- OVCAR3 cells, indicating that miRNAs are involved in the development of this neoplasia and may serve as pertinent chemotherapeutic targets.
癌症干细胞（CSCs）在肿瘤中形成一个非常罕见的群体，具有无限增殖和自我更新的能力。最近已经鉴定出 CD133+卵巢 CSCs（OCSCs），它们的临床意义即将得到阐明。在这种情况下，我们使用 CD133 抗原作为 OVCAR3 细胞中 OCSCs 的标志物，并表明微 RNA（miRNA）在这个亚群中表达异常。使用单克隆鼠抗 CD133-1 抗体（克隆 CD133）鉴定 OVCAR3 细胞系中的 OCSCs。使用微阵列和实时逆转录聚合酶链反应（RT-PCR）分析鉴定 CD133+细胞中表达改变的 miRNA。使用 TaqMan PCR 检查失调 miRNA 的表达水平，即 miR-204、miR-206、miR-223、miR-9、miR-100 和 miR-200c。使用实时 RT-PCR 和 Western blot 分析检查干细胞特异性基因的 RNA 和蛋白质水平。我们的微阵列和实时 RT-PCR 分析结果显示 CD133+和 CD133-OVCAR3 细胞之间存在明显不同的 miRNA 表达谱。CD133+细胞中干细胞特异性基因 Oct3/4、Sox2 和 Nanog 的表达高于 CD133-细胞，而 FoxD3 的表达则低于 CD133-细胞。总之，我们的数据表明，CD133 表达定义了 OVCAR3 细胞系中肿瘤起始细胞的亚群。CD133+OVCAR3 细胞的总体 miRNA 表达谱明显不同于 CD133-OVCAR3 细胞，表明 miRNA 参与了这种肿瘤的发生，并且可能作为相关的化疗靶点。
