METTL14 Promotes Proliferation, Migration, and Invasion in Endometriotic Stromal Cell Growth by Activating the ZEB1/MEK/ERK Pathway.

IF 2 4区 医学 Q2 OBSTETRICS & GYNECOLOGY
Xuan Lv, Fang Li
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引用次数: 0

Abstract

Objective: The aim of this study was to explore the mechanism of methyltransferase-like 14 (METTL14) on human endometriotic stromal cell (ESC; HEM15A) proliferation, migration, and invasion to provide novel therapy for endometriosis (EMs).

Design: Normal human endometrial stromal cells (HESCs) and HEM15A cells were selected. Corresponding controlled experiments were performed to analyze whether overexpression of METTL14, N6-methyladenosine (m6A) methylated ZEB1 mRNA, upregulation of ZEB1, and activating the mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) can affect the proliferation, migration, and invasion of HEM15A cells. Materials, Setting, and Methods: HEM15A and HESCs were cultured in vitro. HEM15A cells were treated with oe-METTL14 and oe-zinc finger E-box-binding protein 1 (ZEB1) plasmids, 3-deazaadenosine (3-DAA) and the MEK/ERK pathway inhibitor isoprenaline (ISO). After identifying HEM15A and HESCs, METTL14, ZEB1, p-ERK1/2/ERK1/2, and p-MEK/MEK levels, and cell proliferation, migration, and invasion were assessed. The modification sites of ZEB1 and m6A were predicted using SRAMP database, with an m6A modification level assessed by MeRIP. The binding of YT521-B homology domain 2 (YTHDF2) to ZEB1 messenger RNA (mRNA), and ZEB1 stability and mRNA level were tested.

Results: Compared with HESCs, METTL14 level in HEM15A was significantly reduced. METTL14 overexpression in HEM15A prominently increased its proliferation, migration, and invasion. METTL14 overexpression notably elevated m6A-methylated ZEB1 mRNA level and reduced the stability and expression of ZEB1 mRNA. Further m6A modification inhibition increased ZEB1 mRNA stability and mRNA and protein levels and decreased ZEB1 m6A modification level. ZEB1 upregulation partially reversed METTL14 overexpression-inhibited HEM15A proliferation, migration, and invasion. METTL14 inhibited the MEK/ERK signaling activation by regulating ZEB1, and the MEK/ERK signaling activation partly averted METTL14-suppressed proliferation, migration, and invasion.

Limitations: The effects of METTL14 on other growth aspects of HEM15A cells and the relation between ZEB1 and m6A require further investigation.

Conclusions: METTL14 lowered ZEB1 expression by regulating ZEB1 m6A modification levels, thereby inhibiting the activation of the MEK/ERK pathway and ESC proliferation, migration, and invasion.

METTL14 通过激活 ZEB1/MEK/ERK 通路,促进子宫内膜异位基质细胞的增殖、迁移和侵袭。
目的:子宫内膜异位症(EMs)常见于生育期妇女。方法:体外培养 HEM15A 和人子宫内膜基质细胞(HESCs)。用oe-METTL14和oe-锌指E-盒结合蛋白1(ZEB1)质粒、N6-甲基腺苷(m6A)抑制剂3-脱氮腺苷(3-DAA)和丝裂原活化蛋白激酶(MEK)/细胞外信号调节激酶(ERK)通路抑制剂异丙肾上腺素(ISO)处理HEM15A细胞。在鉴定 HEM15A 和 HESCs 后,评估了 METTL14、ZEB1、p-ERK1/2/ERK1/2 和 p-MEK/MEK 水平以及细胞增殖、迁移和侵袭情况。利用SRAMP数据库预测了ZEB1和m6A的修饰位点,并通过MeRIP评估了m6A的修饰水平。检测了YT521-B同源结构域2(YTHDF2)与ZEB1信使RNA(mRNA)的结合、ZEB1的稳定性和mRNA水平:结果:与HESCs相比,HEM15A中的METTL14水平明显降低。METTL14在HEM15A中的过表达显著增加了其增殖、迁移和侵袭能力。METTL14 的过表达会明显降低 m6A 甲基化的 ZEB1 mRNA 水平,并降低 ZEB1 mRNA 的稳定性和表达量。进一步的 m6A 修饰抑制增加了 ZEB1 mRNA 的稳定性以及 mRNA 和蛋白质水平,并降低了 ZEB1 m6A 修饰水平。ZEB1 的上调部分逆转了 METTL14 过表达抑制的 HEM15A 增殖、迁移和侵袭。METTL14通过调节ZEB1抑制了MEK/ERK信号的激活,而MEK/ERK信号的激活部分避免了METTL14抑制的增殖、迁移和侵袭:结论:METTL14通过调节ZEB1 m6A修饰水平降低了ZEB1的表达,从而抑制了MEK/ERK通路的激活,抑制了ESC的增殖、迁移和侵袭。
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来源期刊
CiteScore
4.20
自引率
4.80%
发文量
44
审稿时长
6-12 weeks
期刊介绍: This journal covers the most active and promising areas of current research in gynecology and obstetrics. Invited, well-referenced reviews by noted experts keep readers in touch with the general framework and direction of international study. Original papers report selected experimental and clinical investigations in all fields related to gynecology, obstetrics and reproduction. Short communications are published to allow immediate discussion of new data. The international and interdisciplinary character of this periodical provides an avenue to less accessible sources and to worldwide research for investigators and practitioners.
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