Department of Obstetrics and Gynecology, Shanghai First People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, 650 New Songjiang Road, Shanghai 201620, China.
Cancer Cell Int. 2013 May 16;13(1):44. doi: 10.1186/1475-2867-13-44.
Endometrial cancer (EC) is the most common gynecologic malignancy, but the molecular events involved in the development and progression of EC remain unclear. This study aimed to explore epigenetic modification of genes and miRNAs involved in EC development.
Ishikawa and AN3CA cells were treated with 5'-Aza-2-deoxycytidine or histone deacetylase inhibitor. The expression of miRNAs and related genes were detected by PCR and Western blot. Promoter methylation was detected by bisulfite specific PCR sequencing. The proliferation, colony formation, cell cycle progression, migration and invasion of EC cells were evaluated by MTT, soft agar assay, flow cytometry, wound healing and invasion assay, respectively.
Aberrant expression of miRNAs including miR-200b, miR-130a/b, miR-625 and miR-222 was associated with tumorigenesis and metastasis in endometrial cancer. Silencing of miR-130b induced E-cadherin expression, while ectopic expression of miR-130b and knockdown of DICER1 increased the expression of Vimentin, zeb2, N-cadherin, Twist and Snail in EC cells. Furthermore, 5'-Aza-2-deoxycytidine and Histone deacetylase (HDAC) inhibitor inhibited the proliferation, colony formation, migration and invasion of EC cells, accompanied by reduced MMP secretion.
Our study provides the first description of epigenetic modification of epithelial mesenchymal transition associated genes and miRNAs in EC cells, which are extensively involved in the regulation of gene expression and subsequent accumulation of malignant features of EC cells.
子宫内膜癌（EC）是最常见的妇科恶性肿瘤，但涉及 EC 发生和发展的分子事件仍不清楚。本研究旨在探讨参与 EC 发展的基因和 miRNA 的表观遗传修饰。
用 5'-Aza-2-脱氧胞苷或组蛋白去乙酰化酶抑制剂处理 Ishikawa 和 AN3CA 细胞。通过 PCR 和 Western blot 检测 miRNA 和相关基因的表达。通过亚硫酸氢盐特异性 PCR 测序检测启动子甲基化。通过 MTT、软琼脂测定、流式细胞术、划痕愈合和侵袭测定分别评估 EC 细胞的增殖、集落形成、细胞周期进程、迁移和侵袭。
miRNA 的异常表达，包括 miR-200b、miR-130a/b、miR-625 和 miR-222，与子宫内膜癌的发生和转移有关。miR-130b 的沉默诱导 E-钙粘蛋白的表达，而 miR-130b 的异位表达和 DICER1 的敲低增加了 EC 细胞中 Vimentin、zeb2、N-钙粘蛋白、Twist 和 Snail 的表达。此外，5'-Aza-2-脱氧胞苷和组蛋白去乙酰化酶（HDAC）抑制剂抑制 EC 细胞的增殖、集落形成、迁移和侵袭，伴随着 MMP 分泌减少。
本研究首次描述了 EC 细胞中上皮间质转化相关基因和 miRNA 的表观遗传修饰，这些修饰广泛参与了基因表达的调控，随后导致 EC 细胞恶性特征的累积。
