METTL3 介导 hsa_circ_0072380 的 m6A 修饰以调节妊娠期糖尿病的进展。

METTL3 mediates m6A modification of hsa_circ_0072380 to regulate the progression of gestational diabetes mellitus.

机构信息

Department of Obstetrics and Gynecology, The First Affiliated Hospital of Sun Yat-Sen University, Guangdong Provincial Clinical Research Center for Obstetrical and Gynecological Diseases, Guangzhou 510080, China.

Department of Obstetrics and Gynecology, Guangdong Women and Children Hospital, Guangzhou 510010, China.

出版信息

Gene. 2024 Dec 30;931:148894. doi: 10.1016/j.gene.2024.148894. Epub 2024 Aug 25.

DOI:10.1016/j.gene.2024.148894

PMID:39191355

Abstract

BACKGROUND

m6A modification plays a vital role in gestational diabetes mellitus (GDM) progression. However, the role of METTL3 and differential m6A-modified circRNAs in GDMremainsto be investigated.

METHODS

Placental tissue samples from GDM patients and normal controls (NC) were collected to measure changes in m6A modification levels. MeRIP-seq on placental tissue was performed to detect differential m6A-modified circRNAs.High glucose (HG)-treated JEG3 cells were used to establish the GDM cell model. Differentially expressed circRNAs levels in GDM and NC groups were measured by qRT-PCR. We knocked down METTL3 to study its function. Additionally, we conducted functional recovery experiments. Dot blot assay was utilized to assess changes in m6A levels. MeRIP-qPCR was performed to evaluate the effect of knocking down METTL3 on m6A modification of hsa_circ_0072380 in JEG3 cells.

RESULTS

Compared with the NC group, the GDM group exhibited increased levels of m6A modification and METTL3 expression. Differences in m6A modification of circRNAs exist between the GDM and NC groups. Hsa_circ_0000994, hsa_circ_0058733, and hsa_circ_0072380 were significantly down-regulated in the GDM group while hsa_circ_0036376, hsa_circ_0000471, and hsa_circ_0001173 showed no significant differences between two groups. HG treatment promoted METTL3 expression and m6A level of JEG3 cells, and inhibited cell proliferation, migration, and invasion abilities. Knocking down METTL3 reversed these effects. After HG treatment, hsa_circ_0072380 was significantly down-regulated. Knocking down METTL3 led to up-regulation of hsa_circ_0072380, while knocking down hsa_circ_0072380 restored the function of SiMETTL3. Additionally, knocking down METTL3 significantly reduced the m6A modification of hsa_circ_0072380.

CONCLUSION

METTL3 mediated m6A modification of hsa_circ_0072380 to regulate GDM progression.

摘要

背景

m6A 修饰在妊娠糖尿病（GDM）进展中起着至关重要的作用。然而，METTL3 和差异 m6A 修饰的 circRNAs 在 GDM 中的作用仍有待研究。

方法

收集 GDM 患者和正常对照组（NC）的胎盘组织样本，以测量 m6A 修饰水平的变化。对胎盘组织进行 MeRIP-seq，以检测差异表达的 m6A 修饰 circRNAs。用高葡萄糖（HG）处理 JEG3 细胞建立 GDM 细胞模型。通过 qRT-PCR 测量 GDM 和 NC 组之间差异表达 circRNAs 的水平。敲低 METTL3 以研究其功能。此外，我们进行了功能恢复实验。Dot blot 分析用于评估 m6A 水平的变化。MeRIP-qPCR 用于评估敲低 METTL3 对 JEG3 细胞中 hsa_circ_0072380 的 m6A 修饰的影响。

结果

与 NC 组相比，GDM 组的 m6A 修饰和 METTL3 表达水平升高。GDM 和 NC 组之间的 circRNAs 的 m6A 修饰存在差异。hsa_circ_0000994、hsa_circ_0058733 和 hsa_circ_0072380 在 GDM 组中显著下调，而 hsa_circ_0036376、hsa_circ_0000471 和 hsa_circ_0001173 在两组之间无显著差异。HG 处理促进了 JEG3 细胞中 METTL3 的表达和 m6A 水平，抑制了细胞增殖、迁移和侵袭能力。敲低 METTL3 逆转了这些效应。HG 处理后，hsa_circ_0072380 明显下调。敲低 METTL3 导致 hsa_circ_0072380 上调，而敲低 hsa_circ_0072380 恢复了 SiMETTL3 的功能。此外，敲低 METTL3 显著降低了 hsa_circ_0072380 的 m6A 修饰。