Department of Pulmonary, Ningbo No.2 Hospital, Ningbo, China.
Department of Neurology, Ningbo No.2 Hospital, Ningbo, China.
Neurol Res. 2024 Nov;46(11):1083-1093. doi: 10.1080/01616412.2024.2394324. Epub 2024 Aug 29.
Myasthenia gravis (MG) is a both neuromuscular junction and antibody-mediated autoimmune disease, and its pathogenesis involves the regulation of circular RNAs (circRNAs). However, the role of circ_0076490 in MG and the underlying mechanism remain unknown.
RNA levels of circ_0076490, microRNA-144-3p (miR-144-3p), and mitogen-activated protein kinase 1 (MAPK1) were detected by quantitative real-time polymerase chain reaction. Cell viability and proliferation were investigated by cell counting kit-8 and 5-Ethynyl-29-deoxyuridine assays, respectively. Cell cycle and apoptosis were assessed by flow cytometry analysis. The putative binding relationship of miR-144-3p and circ_0076490 or MAPK1 was predicted by circular RNA interactome and TargetScan online databases, respectively, and identified through dual-luciferase reporter assay and RNA pull-down assay.
We observed dramatic increases of circ_0076490 and MAPK1 expression and a decrease of miR-144-3p expression in the peripheral blood of MG patients in comparison with healthy controls. Reduced expression of circ_0076490 induced an inhibitory effect on the proliferation of Jurkat cells and a promoting effect on cell apoptosis. Additionally, miR-144-3p was identified as a target miRNA of circ_0076490, and its depletion attenuated circ_0076490 knockdown-mediated effects on the proliferation and apoptosis of Jurkat cells. MAPK1 was a target gene of miR-144-3p and its overexpression rescued decreased cell proliferation and increased cell apoptosis induced by miR-144-3p introduction. Furthermore, circ_0076490 regulated MAPK1 expression by interacting with miR-144-3p.
Circ_0076490 knockdown inhibited proliferation and induced apoptosis of Jurkat cells through the regulation of the miR-144-3p/MAPK1 axis, providing potential targets for developing improved therapy of MG.
重症肌无力（MG）是一种神经肌肉接头和抗体介导的自身免疫性疾病，其发病机制涉及环状 RNA（circRNA）的调节。然而，circ_0076490 在 MG 中的作用及其潜在机制尚不清楚。
通过实时定量聚合酶链反应检测 circ_0076490、微 RNA-144-3p（miR-144-3p）和丝裂原活化蛋白激酶 1（MAPK1）的 RNA 水平。分别通过细胞计数试剂盒-8 和 5-乙炔基-29-脱氧尿苷测定法检测细胞活力和增殖。通过流式细胞术分析检测细胞周期和凋亡。通过环状 RNA 相互作用组和 TargetScan 在线数据库分别预测 miR-144-3p 与 circ_0076490 或 MAPK1 的假定结合关系，并通过双荧光素酶报告基因检测和 RNA 下拉实验进行鉴定。
与健康对照组相比，MG 患者外周血中 circ_0076490 和 MAPK1 的表达明显增加，miR-144-3p 的表达降低。circ_0076490 表达下调抑制 Jurkat 细胞增殖，促进细胞凋亡。此外，miR-144-3p 被鉴定为 circ_0076490 的靶 miRNA，其耗竭可减弱 circ_0076490 敲低对 Jurkat 细胞增殖和凋亡的影响。MAPK1 是 miR-144-3p 的靶基因，其过表达可挽救 miR-144-3p 引入引起的细胞增殖减少和细胞凋亡增加。此外，circ_0076490 通过与 miR-144-3p 相互作用调节 MAPK1 的表达。
circ_0076490 敲低通过调节 miR-144-3p/MAPK1 轴抑制 Jurkat 细胞增殖并诱导其凋亡，为开发治疗 MG 的新方法提供了潜在靶点。
