雌激素上调DNMT1并导致卵巢子宫内膜异位症恶性转化过程中RUNX3的高甲基化。

Oestrogen up-regulates DNMT1 and leads to the hypermethylation of RUNX3 in the malignant transformation of ovarian endometriosis.

作者信息

Wang Danbo, Guo Cuishan, Li Yan, Zhou Mingyi, Wang Huimin, Liu Jing, Chen Peng

机构信息

Department of Gynecology, Cancer Hospital of China Medical University, Liaoning Cancer Hospital and Institute, Shenyang Liaoning Province 110042, People's Republic of China.

Department of Gynecology and Obstetrics, Shengjing Hospital of China Medical University, Shenyang Liaoning Province 110004, People's Republic of China.

出版信息

Reprod Biomed Online. 2022 Jan;44(1):27-37. doi: 10.1016/j.rbmo.2021.06.030. Epub 2021 Jul 24.

DOI:10.1016/j.rbmo.2021.06.030

PMID:34799276

Abstract

RESEARCH QUESTION

What is the mechanism of hypermethylation of runt-related transcription factor 3 (RUNX3) in the eutopic endometrium of endometriosis as biomarker in the malignant transformation of endometriosis?

DESIGN

Methylation-specific polymerase chain reaction was used to analyse the methylation status of RUNX3 in endometriosis-associated ovarian cancer (EAOC). Primary eutopic endometrial stromal cells (ESC) were isolated from the uteri of patients with ovarian endometriosis. After RUNX3 knockdown by RNA interference technology or ESC treated with oestradiol, the proliferation and invasion ability were evaluated in ESC by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and transwell assays.

RESULTS

The frequency of methylation of RUNX3 in neoplastic tissue in the EAOC group was significantly higher than that in the ectopic endometrium of the endometriosis group (P < 0.001), and the frequency of methylation of RUNX3 in the eutopic endometrium of the EAOC group was significantly higher than that in the endometriosis group (P < 0.001). However, there was no significant difference in the eutopic endometrium when compared between the endometriosis group and the control endometrium group (P = 0.233). Silencing RUNX3 promoted the proliferation and invasion of ESC (P < 0.001 and P < 0.001). Following intervention with oestrogen, it was observed that the oestradiol group showed higher levels of RUNX3 methylation (P < 0.001) and DNA methyltransferase 1 (DNMT1) mRNA and protein expression (P < 0.001 and P < 0.001), and lower RUNX3 mRNA and protein expression when compared with the ESC group (P < 0.001 and P < 0.001).

CONCLUSION

This study demonstrated that hypermethylation of the RUNX3 was related to the malignant transformation of endometriosis and that this process was related to corresponding changes in the eutopic endometrium. Furthermore, the 'oestrogen-DNMT1' signalling pathway may induce the hypermethylation of RUNX3 to promote the malignant transformation of endometriosis.

摘要

研究问题

在子宫内膜异位症的在位内膜中， runt相关转录因子3（RUNX3）高甲基化作为子宫内膜异位症恶变生物标志物的机制是什么？

设计

采用甲基化特异性聚合酶链反应分析子宫内膜异位症相关卵巢癌（EAOC）中RUNX3的甲基化状态。从卵巢子宫内膜异位症患者的子宫中分离出原发性在位内膜基质细胞（ESC）。通过RNA干扰技术敲低RUNX3或用雌二醇处理ESC后，使用3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）和Transwell实验评估ESC中的增殖和侵袭能力。

结果

EAOC组肿瘤组织中RUNX3的甲基化频率显著高于子宫内膜异位症组的异位内膜（P < 0.001），EAOC组在位内膜中RUNX3的甲基化频率显著高于子宫内膜异位症组（P < 0.001）。然而，子宫内膜异位症组与对照内膜组的在位内膜相比无显著差异（P = 0.233）。沉默RUNX3促进了ESC的增殖和侵袭（P < 0.001和P < 0.001）。雌激素干预后，观察到雌二醇组与ESC组相比，RUNX3甲基化水平更高（P < 0.001），DNA甲基转移酶1（DNMT1）mRNA和蛋白表达更高（P < 0.001和P < 0.001），而RUNX3 mRNA和蛋白表达更低（P < 0.001和P < 0.001）。