Identification of miR-30c-5p as a tumor suppressor by targeting the m(6) A reader HNRNPA2B1 in ovarian cancer.
Identification of miR-30c-5p as a tumor suppressor by targeting the m(6) A reader HNRNPA2B1 in ovarian cancer.
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通过靶向卵巢癌中的 m 6 A 阅读器 HNRNPA2B1 鉴定 miR ‐30c 5p 作为肿瘤抑制因子
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microRNA (miRNA) 和 N6-甲基腺苷 (m6A) 在卵巢癌 (OvCa) 中发挥重要作用。然而，迄今为止，miRNA 调节 OvCa 中 m6A 的机制尚未阐明。为了筛选 m6A 相关 miRNA，Pearson 使用癌症基因组图谱数据库 (TCGA) 进行了 miRNA 和 m6A 调节因子的相关性分析。为了确定 m6A 的水平，使用了 RNA m6A 定量测定。然后，进行集落形成测定、EdU 测定、伤口愈合测定和 Transwell 测定。双荧光素酶报告基因检测用于确认 miRNA 靶基因。对目标基因进行蛋白质-蛋白质相互作用（PPI）分析，并使用 cytoHubba/Cytoscape 软件发现枢纽基因。通过生物信息学和 RNA 稳定性测定探索了潜在的分子机制。通过 Pearson 相关性分析，总共有 126 个 miRNA 被鉴定为 m6A 相关 miRNA。其中，高水平的 miR-30c-5p 与 OvCa 患者的良好预后相关。在体外，miR-30c-5p agomir 降低 m6A 水平并抑制 OvCa 细胞增殖、迁移和侵袭。 miR-30c-5p的中心靶基因被确定为（i）XPO1，（ii）AGO1，（iii）HNRNPA2B1，其中m6A阅读器HNRNPA2B1在OvCa组织中高表达，与不良预后相关。在体外，敲低 HNRNPA2B1 显着降低 m6A 水平并阻碍 OvCa 细胞的增殖和迁移。 m6A 阅读器 HNRNPA2B1 的抑制减弱了对增殖和迁移的抑制以及 miR-30c-5p 下调引起的低 m6A 水平。从机制上讲，m6A 阅读器 HNRNPA2B1 可能通过调节 CDK19 mRNA 稳定性来改变 m6A 水平。 miR-30c-5p 通过抑制 m6A reader HNRNPA2B1 抑制 OvCa 进展并降低 m6A 水平，从而为 OvCa 中 m6A 调控机制提供新的见解。 miR-30c-5p 通过抑制 m6A reader HNRNPA2B 来抑制 OvCa 进展。同时，HNRNPA2B1可能调节CDK19 mRNA稳定性以改变m6A水平。
microRNAs (miRNAs) and N6‐methyladenosine (m6A) play important roles in ovarian cancer (OvCa). However, the mechanisms by which miRNAs regulate m6A in OvCa have not been elucidated so far. To screen m6A‐related miRNAs, Pearson's correlation analysis of miRNAs and m6A regulators was implemented using The Cancer Genome Atlas database (TCGA). To determine the level of m6A, RNA m6A quantitative assays were used. Then, colony formation assays, EdU assays, wound healing assays, and Transwell assays were performed. The dual‐luciferase reporter assay was used to confirm the miRNA target genes. Protein–protein interaction (PPI) analysis of the target genes was performed, and hub genes were discovered using the cytoHubba/Cytoscape software. The underlying molecular mechanisms were explored by bioinformatics and RNA stability assays. A total of 126 miRNAs were identified as m6A‐related miRNAs by Pearson's correlation analysis. Among them, the high level of miR‐30c‐5p was associated with good prognosis in OvCa patients. In vitro, the miR‐30c‐5p agomir lowered the m6A level and inhibited OvCa cell proliferation, migration, and invasion. The hub target genes of miR‐30c‐5p were identified as (i) XPO1, (ii) AGO1, (iii) HNRNPA2B1, of which m6A reader HNRNPA2B1 was highly expressed in OvCa tissues and related with poor prognosis. In vitro, knockdown of HNRNPA2B1 significantly reduced m6A level and hampered the proliferation and migration of OvCa cells. The inhibition of m6A reader HNRNPA2B1 attenuated the suppression of proliferation and migration and the low m6A level induced by the miR‐30c‐5p downregulation. Mechanistically, m6A reader HNRNPA2B1 might regulate CDK19 mRNA stability to alter m6A level. miR‐30c‐5p inhibits OvCa progression and reduces the m6A level by inhibiting m6A reader HNRNPA2B1, thus providing new insights into the m6A regulatory mechanism in OvCa. miR‐30c‐5p inhibits OvCa progression by inhibiting m6A reader HNRNPA2B. Meanwhile, HNRNPA2B1 might regulate CDK19 mRNA stability to alter m6A level.