Key Laboratory of Assisted Reproduction, Ministry of Education, Center of Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100191, PR China.
Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, PR China.
Gynecol Oncol. 2017 Nov;147(2):408-417. doi: 10.1016/j.ygyno.2017.08.010. Epub 2017 Aug 26.
Epithelial ovarian cancer (EOC) is the deadliest type of ovarian cancer, but the mechanisms contributing to its tumorigenesis are not well understood. Herein, we will elucidate the role of Ten-eleven translocation 1 (TET1) in EOC development.
The expression of TET1 in EOC cell lines and primary samples was examined by western blot and immunohistochemistry. The biological role of ectopic TET1 overexpression was revealed by a series of in vitro functional studies. Its downstream signaling pathway was predicted by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of microarray data. The methylation level and expression of Wnt/β-catenin signaling inhibitors Dikkopf 1 (DKK1) and secreted Fzd receptor protein 2 (SFRP2) were examined by Chromatin immunoprecipitation (ChIP) assay, Epimark™ 5hmC and 5mC level analysis and quantitative RT-PCR. Small interference RNA (siRNA) technology was used to investigate the biological roles of DKK1 and SFRP2.
TET1 expression was inversely correlated with clinical stage in patients with EOC by tissue microarray (TMA). TET1 expression was undetected in 6 types of EOC cell lines. Ectopic expression of TET1 inhibited colony formation, cell migration and invasion in SKOV3 and OVCAR3 cells. Furthermore, TET1 overexpression reversed the epithelial-mesenchymal transition (EMT) process of SKOV3 cells. Mechanistically, TET1 potently inhibited canonical Wnt/β-catenin signaling by demethylating and upregulating two upstream antagonists of this pathway, SFRP2 and DKK1, which was associated with inhibition of EMT and cancer cell metastasis.
This study uncovers that TET1 has potent tumor-suppressive effects in EOC by activating Wnt/β-catenin signaling inhibitors DKK1 and SFRP2.
上皮性卵巢癌(EOC)是卵巢癌中致死率最高的一种,但导致其发生的机制尚不清楚。在此,我们将阐明 Ten-eleven translocation 1(TET1)在 EOC 发展中的作用。
通过 Western blot 和免疫组织化学检测 TET1 在 EOC 细胞系和原代样本中的表达。通过一系列体外功能研究揭示了异位 TET1 过表达的生物学作用。通过微阵列数据的京都基因与基因组百科全书(KEGG)通路分析预测其下游信号通路。通过染色质免疫沉淀(ChIP)检测、Epimark™ 5hmC 和 5mC 水平分析和定量 RT-PCR 检测 Wnt/β-catenin 信号抑制剂 Dikkopf 1(DKK1)和分泌型 Fzd 受体蛋白 2(SFRP2)的甲基化水平和表达。使用小干扰 RNA(siRNA)技术研究 DKK1 和 SFRP2 的生物学作用。
通过组织微阵列(TMA)发现 TET1 表达与 EOC 患者的临床分期呈负相关。6 种 EOC 细胞系中未检测到 TET1 表达。TET1 过表达抑制 SKOV3 和 OVCAR3 细胞的集落形成、细胞迁移和侵袭。此外,TET1 过表达逆转了 SKOV3 细胞的上皮-间充质转化(EMT)过程。机制上,TET1 通过去甲基化和上调该通路的两个上游拮抗剂 SFRP2 和 DKK1,强力抑制了经典的 Wnt/β-catenin 信号,这与 EMT 和癌细胞转移的抑制有关。
本研究揭示 TET1 通过激活 Wnt/β-catenin 信号抑制剂 DKK1 和 SFRP2,在上皮性卵巢癌中具有强大的肿瘤抑制作用。