Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal human cancers. It thrives in a malnourished environment; however, little is known about the mechanisms by which PDAC cells actively promote aerobic glycolysis to maintain their metabolic needs. Gene Expression Omnibus (GEO) was used to identify differentially expressed miRNAs. The expression pattern of miR-30d in normal and PDAC tissues was studied by in situ hybridization. The role of miR-30d/RUNX1 in vitro and in vivo was evaluated by CCK8 assay and clonogenic formation as well as transwell experiment, subcutaneous xenograft model and liver metastasis model, respectively. Glucose uptake, ATP and lactate production were tested to study the regulatory effect of miR-30d/RUNX1 on aerobic glycolysis in PDAC cells. Quantitative real-time PCR, western blot, Chip assay, promoter luciferase activity, RIP, MeRIP, and RNA stability assay were used to explore the molecular mechanism of YTHDC1/miR-30d/RUNX1 in PDAC. Here, we discover that miR-30d expression was remarkably decreased in PDAC tissues and associated with good prognosis, contributed to the suppression of tumor growth and metastasis, and attenuation of Warburg effect. Mechanistically, the m⁶A reader YTHDC1 facilitated the biogenesis of mature miR-30d via m⁶A-mediated regulation of mRNA stability. Then, miR-30d inhibited aerobic glycolysis through regulating SLC2A1 and HK1 expression by directly targeting the transcription factor RUNX1, which bound to the promoters of the SLC2A1 and HK1 genes. Moreover, miR-30d was clinically inversely correlated with RUNX1, SLC2A1 and HK1, which function as adverse prognosis factors for overall survival in PDAC tissues. Overall, we demonstrated that miR-30d is a functional and clinical tumor-suppressive gene in PDAC. Our findings further uncover that miR-30d is a novel target for YTHDC1 through m⁶A modification, and miR-30d represses pancreatic tumorigenesis via suppressing aerobic glycolysis.

胰腺导管腺癌 (PDAC) 是最致命的人类癌症之一。它在营养不良的环境中茁壮成长；然而，对于 PDAC 细胞积极促进有氧糖酵解以维持其代谢需求的机制知之甚少。Gene Expression Omnibus (GEO) 用于鉴定差异表达的 miRNA。通过原位杂交研究miR-30d在正常组织和PDAC组织中的表达模式。miR-30d/RUNX1在体外和体内的作用分别通过CCK8测定和克隆形成以及transwell实验、皮下异种移植模型和肝转移模型进行评估。测试葡萄糖摄取、ATP 和乳酸生成以研究 miR-30d/RUNX1 对 PDAC 细胞有氧糖酵解的调节作用。定量实时 PCR、蛋白质印迹、芯片检测、启动子荧光素酶活性、RIP、MeRIP 和 RNA 稳定性测定用于探索 YTHDC1/miR-30d/RUNX1 在 PDAC 中的分子机制。在这里，我们发现 PDAC 组织中 miR-30d 表达显着降低并与良好预后相关，有助于抑制肿瘤生长和转移，并减弱 Warburg 效应。从机械上讲，m⁶阅读器 YTHDC1 通过 m ⁶ A 介导的 mRNA 稳定性调节促进成熟 miR-30d 的生物发生。然后，miR-30d 通过直接靶向与 SLC2A1 和 HK1 基因的启动子结合的转录因子 RUNX1 来调节 SLC2A1 和 HK1 的表达，从而抑制有氧糖酵解。此外，miR-30d 在临床上与 RUNX1、SLC2A1 和 HK1 呈负相关，它们是 PDAC 组织中总体生存的不良预后因素。总的来说，我们证明了 miR-30d 是 PDAC 中的功能性和临床肿瘤抑制基因。我们的研究结果进一步表明，miR-30d 是 YTHDC1 通过 m ⁶ A 修饰的新靶标，并且 miR-30d 通过抑制有氧糖酵解来抑制胰腺肿瘤发生。