葡萄糖参与 2 型糖尿病患者 m6A 的动态调控。

Glucose Is Involved in the Dynamic Regulation of m6A in Patients With Type 2 Diabetes.

机构信息

Center for Gene Diagnosis, Zhongnan Hospital of Wuhan University, Wuhan, China.

College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, China.

出版信息

J Clin Endocrinol Metab. 2019 Mar 1;104(3):665-673. doi: 10.1210/jc.2018-00619.

DOI:10.1210/jc.2018-00619

PMID:30137347

Abstract

CONTEXT

N6-methyladenosine (m6A) in mRNA is the most abundant and reversible modification. However, the mechanism behind the decrease in m6A in patients with type 2 diabetes (T2D) has not yet been thoroughly investigated.

OBJECTIVE

To clarify whether glucose is involved in the dynamic regulation of m6A in T2D and to identify a possible underlying mechanism.

METHODS

Liquid chromatography/electrospray ionization/tandem mass spectrometry and quantitative PCR were performed to determine the m6A content and the mRNA expression of target genes in 102 patients with T2D and 107 controls. An additional 12 patients with normal fasting blood glucose, emergency hyperglycemia, or emergency hypoglycemia, as well as HepG2 cells with high-glucose treatment and FTO knockout or overexpression were used to confirm the initial observations in patients.

RESULTS

In patients with T2D, the m6A content was decreased, and mRNA expression levels of FTO, METTL3, METTL14, and WTAP were increased. Interestingly, the m6A content was negatively associated with mRNA expression levels of METTL3, METTL14, and FTO. Moreover, FTO was positively correlated with serum glucose. In HepG2 cells, high glucose upregulated FTO protein, whereas it had no significant effect on METTL3 or METTL14. Additionally, mRNA expression levels of FOXO1, G6PC, and DGAT2 were significantly increased and positively correlated with FTO and serum glucose in patients.

CONCLUSIONS

Our data revealed that in patients with T2D, high-glucose-enhanced FTO mRNA expression resulted in a decrease in m6A. The lower m6A content might be responsible for the upregulation of methyltransferases. Additionally, FTO induced mRNA expression of FOXO1, G6PC, and DGAT2 and was closely associated with glucose metabolism.

摘要

背景

mRNA 中的 N6-甲基腺苷（m6A）是最丰富和最可逆转的修饰。然而，2 型糖尿病（T2D）患者中 m6A 减少的机制尚未得到深入研究。

目的

阐明葡萄糖是否参与 T2D 中 m6A 的动态调节，并确定潜在的机制。

方法

采用液相色谱/电喷雾电离/串联质谱和定量 PCR 检测 102 例 T2D 患者和 107 例对照者的 m6A 含量和靶基因的 mRNA 表达。另外，对 12 例空腹血糖正常、应急性高血糖或应急性低血糖的患者以及高糖处理的 HepG2 细胞和 FTO 敲除或过表达的 HepG2 细胞进行验证。

结果

在 T2D 患者中，m6A 含量降低，FTO、METTL3、METTL14 和 WTAP 的 mRNA 表达水平升高。有趣的是，m6A 含量与 METTL3、METTL14 和 FTO 的 mRNA 表达水平呈负相关。此外，FTO 与血清葡萄糖呈正相关。在 HepG2 细胞中，高糖上调了 FTO 蛋白，但对 METTL3 或 METTL14 没有显著影响。此外，患者中 FOXO1、G6PC 和 DGAT2 的 mRNA 表达水平显著增加，与 FTO 和血清葡萄糖呈正相关。