胰腺癌（PC）是恶性程度最高的癌症之一，具有高度侵袭性和预后不良的特点。 N6-甲基腺苷 (m6A) 已被证明参与 PC 的开发。葡聚糖分支酶1（GBE1）主要参与细胞糖原代谢。然而，GBE1的功能以及GBE1在PC进展中是否发生m6A修饰仍有待阐明。通过在线平台分析GBE1的临床预后。 GBE1的表达是从在线平台获得的，然后在正常细胞系和PC细胞系中进行验证。慢病毒用于生成 GBE1 稳定过表达或敲低的 PC 细胞。采用细胞计数试剂盒（CCK-8）、集落形成实验、球体形成实验和流式细胞仪实验分析细胞体外增殖和干细胞能力。使用皮下和原位小鼠模型来验证GBE1的体内功能。通过RNA免疫沉淀（RIP）实验、RNA稳定性实验和蛋白质印迹来探讨GBE1在PC中的分子调控。GBE1在PC中显着上调，并与PC患者的不良预后相关。从功能上来说，GBE1的过表达促进了PC细胞的增殖和干细胞样特性，而GBE1的敲低则减弱了PC细胞的恶性程度。重要的是，我们发现 GBE1 RNA 的 m6A 修饰，并且 WTAP 和 IGF2BP3 被揭示为 m6A 调节因子，可以增加 GBE1 mRNA 的稳定性和表达。此外，c-Myc 被发现是 GBE1 的下游基因，功能拯救实验表明 c-Myc 的过表达可以拯救 GBE1 敲低诱导的 PC 细胞生长抑制。我们的研究揭示了 GBE1/c-Myc 轴在 PC 中的致癌作用进展并揭示了 WTAP/IGF2BP3 介导的 GBE1 m6A 修饰，这突出了 GBE1 在 PC 靶向治疗中的潜在应用。© 2024。作者。

Pancreatic cancer (PC) is one of the most malignant cancers with highly aggressiveness and poor prognosis. N6-methyladenosine (m6A) have been indicated to be involved in PC development. Glucan Branching Enzyme 1 (GBE1) is mainly involved in cell glycogen metabolism. However, the function of GBE1 and Whether GBE1 occurs m6A modification in PC progression remains to be illustrated.The clinical prognosis of GBE1 was analyzed through online platform. The expression of GBE1 was obtained from online platform and then verified in normal and PC cell lines. Lentivirus was used to generated GBE1 stable-overexpression or knockdown PC cells. Cell Counting Kit (CCK-8), colony formation assay, sphere formation assay and flow cytometry assay were conducted to analyze cell proliferation and stemness ability in vitro. Subcutaneous and orthotopic mouse models were used to verify the function of GBE1 in vivo. RNA immunoprecipitation (RIP) assay, RNA stability experiment and western blots were conducted to explore the molecular regulation of GBE1 in PC.GBE1 was significantly upregulated in PC and associated with poor prognosis of PC patients. Functionally, GBE1 overexpression facilitated PC cell proliferation and stemness-like properties, while knockdown of GBE1 attenuated the malignancy of PC cells. Importantly, we found the m6A modification of GBE1 RNA, and WTAP and IGF2BP3 was revealed as the m6A regulators to increase GBE1 mRNA stability and expression. Furthermore, c-Myc was discovered as a downstream gene of GBE1 and functional rescue experiments showed that overexpression of c-Myc could rescue GBE1 knockdown-induced PC cell growth inhibition.Our study uncovered the oncogenic role of GBE1/c-Myc axis in PC progression and revealed WTAP/IGF2BP3-mediated m6A modification of GBE1, which highlight the potential application of GBE1 in the targeted therapy of PC.© 2024. The Author(s).