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FTO 敲低介导的 miR-383-5p 成熟通过靶向 ITGA3 抑制胰腺癌细胞的恶性进展。

FTO Knockdown-Mediated Maturation of miR-383-5p Inhibits Malignant Advancement of Pancreatic Cancer by Targeting ITGA3.

机构信息

Department of Radiology, Children's Hospital of Nanjing Medical University, No. 72, Guangzhou Road, Gulou District, Nanjing, 210008, China.

Department of Interventional and Vascular Surgery, Shanghai Tenth People's Hospital, Tongji University School of Medicine, No. 301, Yanchang Road, Jing'an District, Shanghai, 200072, China.

出版信息

Biochem Genet. 2024 Aug;62(4):2652-2666. doi: 10.1007/s10528-023-10560-0. Epub 2023 Nov 24.

Abstract

m6A demethylase FTO is confirmed to be involved in pancreatic cancer progression. FTO regulates miRNA processing. To investigate the regulatory effect of FTO on miR-383-5p and its role in pancreatic cancer. The expression of miR-383-5p, ITGA3, and FTO was predicted using bioinformatic analysis in tissues and was measured using qPCR in cells. Cell biological functions were investigated using MTT assay, Transwell assay, sphere formation assay, and qPCR. The targeting relationship between miR-383-5p and ITGA3 was evaluated using the dual-luciferase reporter assay. The effect of FTO on miR-383-5p processing was evaluated using RIP and MeRIP assay. FTO expression was upregulated in pancreatic cancer and silencing of FTO promoted the processing of miR-383-5p in an m6A-dependent manner. m6A-modified miRNA processing was recognized by IGF2BP1. Downregulation of miR-383-5p reversed FTO knockdown-induced inhibition of cellular processes. The FTO/miR-383-5p/ITGA3 axis facilitated cell viability, metastasis, and stemness in pancreatic cancer.

摘要

m6A 去甲基酶 FTO 被证实参与胰腺癌的进展。FTO 调节 miRNA 的加工。为了研究 FTO 对 miR-383-5p 的调控作用及其在胰腺癌中的作用。采用生物信息学分析预测组织中 miR-383-5p、ITGA3 和 FTO 的表达,并在细胞中通过 qPCR 进行测量。通过 MTT 测定、Transwell 测定、球体形成测定和 qPCR 研究细胞生物学功能。通过双荧光素酶报告基因测定评估 miR-383-5p 和 ITGA3 之间的靶向关系。通过 RIP 和 MeRIP 测定评估 FTO 对 miR-383-5p 加工的影响。FTO 在胰腺癌中表达上调,沉默 FTO 以 m6A 依赖性方式促进 miR-383-5p 的加工。m6A 修饰的 miRNA 加工被 IGF2BP1 识别。下调 miR-383-5p 逆转了 FTO 敲低诱导的细胞过程抑制。FTO/miR-383-5p/ITGA3 轴促进了胰腺癌中的细胞活力、转移和干性。

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