文章：

Hsa_circ_0007590/PTBP1复合物通过降低m6A修饰的PTEN mRNA的稳定性，重编程胰腺导管腺癌中的葡萄糖代谢

Hsa_circ_0007590/PTBP1 complex reprograms glucose metabolism by reducing the stability of m6A-modified PTEN mRNA in pancreatic ductal adenocarcinoma

原文发布日期：2024-05-27 

英文摘要：

The role of circular RNAs (circRNAs) in glucose metabolism in pancreatic duct adenocarcinoma (PDAC) remains elusive. Through RNA sequencing of cells cultured under conditions of glucose deprivation, we identified hsa_circ_0007590. Sanger sequencing and RNase R and Act D treatments were performed to confirm the circular RNA features of hsa_circ_0007590. RNA in situ hybridization (RNA-ISH) and quantitative reverse transcription PCR (qRT-PCR) were used to estimate hsa_circ_0007590 expression in PDAC clinical specimens and cell lines. hsa_circ_0007590 expression was higher in PDAC patients and closely related to the clinicopathological characteristics of the disease. Cytoplasm‒nuclear fractionation and FISH assays demonstrated that hsa_circ_0007590 was located in the nucleus. Gain-of-function and loss-of-function assays were performed to assess the biological behaviors of PDAC cells. Seahorse XF assays were performed to validate the Warburg effect. hsa_circ_0007590 facilitated the proliferation, migration, and invasion of PDAC cells and promoted the Warburg effect. Mass spectrometry, RNA pulldown, RNA immunoprecipitation (RIP), RNA m6A quantification, m6A dot blot, MeRIP, and Western blotting were conducted to investigate the detailed mechanism through which hsa_circ_0007590 produces these effects. Mechanistically, hsa_circ_0007590 targeted PTBP1 and increased the expression of the m6A reader protein YTHDF2, leading to PTEN mRNA degradation and PI3K/AKT/mTOR pathway activation. Overall, hsa_circ_0007590, which targets PTBP1, reprograms glucose metabolism by attenuating the stability of m6A-modified PTEN mRNA and holds potential promise as a therapeutic target for PDAC.

摘要翻译：

环状RNA（circRNA）在胰腺导管腺癌（PDAC）葡萄糖代谢中的作用尚不明确。通过葡萄糖剥夺培养条件下的细胞RNA测序，我们鉴定出hsa_circ_0007590。经Sanger测序、RNase R和放线菌素D处理证实其环状RNA特性。采用RNA原位杂交（RNA-ISH）和定量逆转录PCR（qRT-PCR）检测PDAC临床标本和细胞系中hsa_circ_0007590的表达水平，发现其在PDAC患者中表达升高且与临床病理特征密切相关。细胞质-核分离及FISH实验表明hsa_circ_0007590定位于细胞核。功能获得与缺失实验显示：hsa_circ_0007590可促进PDAC细胞增殖、迁移和侵袭，并通过Seahorse XF分析验证其增强Warburg效应。通过质谱分析、RNA pulldown、RNA免疫沉淀（RIP）、RNA m6A定量、m6A斑点印迹、MeRIP和Western blotting等技术深入探讨其作用机制，发现hsa_circ_0007590靶向PTBP1并增加m6A阅读蛋白YTHDF2的表达，导致PTEN mRNA降解及PI3K/AKT/mTOR通路激活。综上，hsa_circ_0007590通过靶向PTBP1削弱m6A修饰的PTEN mRNA稳定性，重编程葡萄糖代谢过程，有望成为PDAC的治疗靶点。

原文链接：

Hsa_circ_0007590/PTBP1 complex reprograms glucose metabolism by reducing the stability of m6A-modified PTEN mRNA in pancreatic ductal adenocarcinoma