Estrogen-mediated DNMT1 and DNMT3A recruitment by EZH2 silences miR-570-3p that contributes to papillary thyroid malignancy through DPP4.

IF 4.8 2区医学 Q1 GENETICS & HEREDITY

Clinical Epigenetics Pub Date : 2024-06-18 DOI:10.1186/s13148-024-01685-z

Xiarong Hu, Qingyao Ye, HuanQuan Lu, Zhiming Wu, Siyuan Chen, Ruinian Zheng

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引用次数: 0

Abstract

Background: Papillary thyroid carcinoma (PTC) is a common endocrine malignancy. Studies have indicated that estrogen can regulate the expression of miRNAs in numerous malignancies. MiR-570-3p has been shown to have a regulatory function in various cancers. However, studies of the regulatory function of miR-570-3p and a direct link between estrogen (especially estradiol E2) and miR-570-3p in PTC have not been done.

Methods: Expression of miR-570-3p and its downstream target DPP4 in PTC tissues and cells was predicted using bioinformatics and validated by qRT-PCR and western blot assays. We then performed a series of gain-and-loss experiments to assess the functional significance of miR-570-3p/DPP4 axis in PTC progression in vitro and in vivo. Additionally, the methylation of the miR-570-3p promoter region was examined via bioinformatics analysis and MSP. Finally, the effects of E2 on PTC progression and the correlation between DNMT1/DNMT3A and EZH2 were predicted by bioinformatic tools and proved by luciferase reporter, ChIP, and co-IP assays.

Results: In PTC tumor tissues and cell lines, there was a lower expression level and a higher methylation level of miR-570-3p compared to normal tissues and cell lines. DPP4 was identified as the downstream target of miR-570-3p. Overexpression of miR-570-3p reduced the proliferative, migratory, and invasive capabilities, and promoted apoptosis, while overexpression of DPP4 reversed these effects in PTC cells. It was also discovered that DNMT1 and DNMT3A increased the CpG methylation level of the miR-570-3p promoter in an EZH2-dependent manner, which led to decreased expression of miR-570-3p. Furthermore, we observed that estrogen (E2) enhanced the methylation of miR-570-3p and suppressed its expression levels, resulting in augmented tumor growth in vivo in PTC.

Conclusion: Estrogen regulates the EZH2/DNMTs/miR-570-3p/DPP4 signaling pathway to promote PTC progression.

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EZH2 通过雌激素介导的 DNMT1 和 DNMT3A 招募抑制了 miR-570-3p，而 miR-570-3p 则通过 DPP4 导致甲状腺乳头状恶性肿瘤。

背景：甲状腺乳头状癌（PTC）是一种常见的内分泌恶性肿瘤：甲状腺乳头状癌（PTC）是一种常见的内分泌恶性肿瘤。研究表明，雌激素能调节多种恶性肿瘤中 miRNAs 的表达。MiR-570-3p 已被证明在多种癌症中具有调控功能。然而，关于miR-570-3p的调控功能以及雌激素（尤其是雌二醇E2）和miR-570-3p在PTC中的直接联系的研究尚未完成：方法：利用生物信息学方法预测了 miR-570-3p 及其下游靶点 DPP4 在 PTC 组织和细胞中的表达，并通过 qRT-PCR 和 Western 印迹检测进行了验证。然后，我们进行了一系列增减实验，以评估 miR-570-3p/DPP4 轴在体外和体内 PTC 进展中的功能意义。此外，还通过生物信息学分析和 MSP 研究了 miR-570-3p 启动子区域的甲基化情况。最后，通过生物信息学工具预测了E2对PTC进展的影响以及DNMT1/DNMT3A和EZH2之间的相关性，并通过荧光素酶报告、ChIP和co-IP检测进行了证实：结果：在 PTC 肿瘤组织和细胞系中，与正常组织和细胞系相比，miR-570-3p 的表达水平较低，甲基化水平较高。DPP4被确定为miR-570-3p的下游靶标。过表达 miR-570-3p 可降低 PTC 细胞的增殖、迁移和侵袭能力，促进细胞凋亡，而过表达 DPP4 则可逆转这些效应。研究还发现，DNMT1 和 DNMT3A 以 EZH2 依赖性的方式增加了 miR-570-3p 启动子的 CpG 甲基化水平，从而导致 miR-570-3p 的表达下降。此外，我们还观察到雌激素（E2）增强了miR-570-3p的甲基化，抑制了其表达水平，从而导致PTC体内肿瘤生长增强：结论：雌激素调节 EZH2/DNMTs/miR-570-3p/DPP4 信号通路，促进 PTC 的进展。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Clinical Epigenetics ONCOLOGY-

自引率

5.30%

发文量

150

期刊介绍： Clinical Epigenetics, the official journal of the Clinical Epigenetics Society, is an open access, peer-reviewed journal that encompasses all aspects of epigenetic principles and mechanisms in relation to human disease, diagnosis and therapy. Clinical trials and research in disease model organisms are particularly welcome.