Abstract GMM-017: UPREGULATION OF MIR-328 CONTRIBUTES TO OVARIAN CANCER STEM CELL MAINTENANCE BY DOWNREGULATING DDB2

Abstract

Abstract Cancer stem cells (CSCs) are a particular subpopulation of cells that are characterized by self renewal, differentiation and enhanced tumorigenicity. They are responsible for tumor metastasis, relapse and development of drug resistance. Thus, eradication of CSCs is essential for improved patient prognosis. Micro RNAs are a group of small non-coding, endogenous RNAs that are found to regulate cancer stem cell characteristics by binding to mRNA in a sequence specific manner. In ovarian cancers, a wide array of Micro RNAs have been found to show differential expression of which miR328-3p deserves special mention. In this study, a Micro RNA Nanostring profile analysis reveals a significant upregulation of miR-328-3p in ovarian cancer stem cells isolated from both ovarian cancer cell lines and primary ovarian tumors as compared to their corresponding bulk cells. Moreover, it was found that inhibition of miR-328 limited the CSC population in ovarian cancer cells whereas overexpression of miR-328 enriched the CSC population, thus accounting for miR-328 as an onco-miRNA. The upregulation of miR-328 not only increased the percentage of ALDH+ cells in ovarian cancer bulk cells, but also increased the tumorigenicity and sphere formation ability. This was supported by the orthotopic ovarian xenograft assay. Further investigation revealed that reduced phosphorylation of Erk in ovarian cancer stem cells owing to reduced levels of Reactive Oxygen species (ROS) could be a prospective mechanism behind elevated miR328 expression and maintenance of CSC characteristics. Inhibition of phosphorylated Erk expression in ovarian cancer bulk cells by use of commercially available Erk inhibitor, U0126, led to a significant increase in miR328 expression. Simultaneously, upregulation of phosphorylated Erk in ovarian cancer stem cells not only reduced miR328 expression, but also displayed a significant reduction in expression of cancer stem cell markers (Oct4, Sox2, Nanog), sphere formation ability and tumorigenesis. We obtained a similar trend of results on regulating the expression of pErk by use of Reactive Oxygen Species to ovarian cancer cells. These data further helped us confirm our speculation that reduced ROS promotes the maintenance of CSCs characteristics through inactivation of Erk signalling pathway. Besides, we also identified DDB2 as a direct target of miR328. Our previous findings demonstrate that DDB2 is able to limit ovarian CSC population by disrupting their self renewal capacity. Thus, we conclude that elevated miR328 in ovarian CSCs, resulting from inactivated Erk1/2 activity, is responsible for maintenance of stemness by inhibition of DDB2 expression. Targeting miR-328 could therefore be a novel therapeutic strategy to eradicate CSCs in ovarian cancer. Citation Format: Amit Kumar Srivastava, Tiantian Cui, Ananya Banerjee, Chunhua Han, Shurui Cai, Lu Liu, Dayong Wu1, Ri Cui, Zaibo Li, Xiaoli Zhang, Guozhen Xie, Selvendiran Karuppalyah, Adam Karpf, Jinsong Liu, David Cohn, Qi-En Wang. UPREGULATION OF MIR-328 CONTRIBUTES TO OVARIAN CANCER STEM CELL MAINTENANCE BY DOWNREGULATING DDB2 [abstract]. In: Proceedings of the 12th Biennial Ovarian Cancer Research Symposium; Sep 13-15, 2018; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2019;25(22 Suppl):Abstract nr GMM-017.