miR-137 is a tumor suppressor in endometrial cancer and is repressed by DNA hypermethylation

Endometrial cancer is the most common gynecological cancer in the United States. We wanted to identify epigenetic aberrations involving microRNAs (miRNAs), whose genes become hypermethylated in endometrial primary tumors. By integrating known miRNA sequences from the miRNA database (miRBase) with DNA methylation data from methyl-CpG-capture sequencing, we identified 111 differentially methylated regions (DMRs) associated with CpG islands (CGIs) and miRNAs. Among them, 22 DMRs related to 29 miRNAs and within 8 kb of CGIs were hypermethylated in endometrial tumors but not in normal endometrium. miR-137was further validated in additional endometrial primary tumors. Hypermethylation of miR-137was found in both endometrioid and serous endometrial cancer (P< 0.01), and it led to the loss of miR-137expression. Treating hypermethylated endometrial cancer cells with epigenetic inhibitors reactivated miR-137. Moreover, genetic overexpression of miR-137suppressed cancer cell proliferation and colony formation in vitro. When transfected cancer cells were implanted into nude mice, the cells that overexpressed miR-137grew more slowly and formed smaller tumors (P< 0.05) than vector transfectants. Histologically, xenograft tumors from cancer cells expressing miR-137were less proliferative (P< 0.05), partly due to inhibition of EZH2 and LSD1 expression (P< 0.01) in both the transfected cancer cells and tumors. Reporter assays indicated that miR-137targets EZH2 and LSD1. These results suggest that miR-137is a tumor suppressor that is repressed in endometrial cancer because the promoter of its gene becomes hypermethylated.