Table_3_The m6A methyltransferase METTL16 inhibits the proliferation of pancreatic adenocarcinoma cancer cells via the p21 signaling pathway.xlsx

BackgroundMany studies have reported that N6-methyladenosine (m6A) modification plays a critical role in the epigenetic regulation of organisms and especially in the pathogenesis of malignant diseases. However, m6A research has mainly focused on methyltransferase activity mediated by METTL3, and few studies have focused on METTL16. The aim of this study was to investigate the mechanism of METTL16, which mediates m6A modification, and its role in pancreatic adenocarcinoma (PDAC) cell proliferation.MethodsClinicopathologic and survival data were retrospectively collected from 175 PDAC patients from multiple clinical centers to detect the expression of METTL16. CCK-8, cell cycle, EdU and xenograft mouse model experiments were used to evaluate the proliferation effect of METTL16. Potential downstream pathways and mechanisms were explored via RNA sequencing, m6A sequencing, and bioinformatic analyses. Regulatory mechanisms were studied through methyltransferase inhibition, RIP, MeRIP‒qPCR assays.ResultsWe found that METTL16 expression was markedly downregulated in PDAC, and multivariate Cox regression analyses revealed that METTL16 was a protective factor for PDAC patients. We also demonstrated that METTL16 overexpression inhibited PDAC cell proliferation. Furthermore, we identified a METTL16-p21 signaling axis, with downregulation of METTL16 resulting in inhibition of CDKN1A (p21). Additionally, METTL16 silencing and overexpression experiments highlighted m6A modification alterations in PDAC.ConclusionsMETTL16 plays a tumor-suppressive role and suppresses PDAC cell proliferation through the p21 pathway by mediating m6A modification. METTL16 may be a novel marker of PDAC carcinogenesis and target for the treatment of PDAC.

背景：众多研究指出，N6-甲基腺苷（m6A）修饰在生物体的表观遗传调控中扮演着至关重要的角色，尤其在恶性肿瘤的发病机制中尤为重要。然而，m6A研究主要集中在由METTL3介导的甲基转移酶活性上，而针对METTL16的研究则相对较少。本研究旨在探究介导m6A修饰的METTL16的机制及其在胰腺腺癌（PDAC）细胞增殖中的作用。方法：通过回顾性收集来自多个临床中心的175例PDAC患者的临床病理和生存数据，检测METTL16的表达。采用CCK-8、细胞周期、EdU和异种移植小鼠模型实验评估METTL16对增殖的影响。通过RNA测序、m6A测序和生物信息学分析探索潜在的下游途径和机制。通过甲基转移酶抑制、RIP、MeRIP-qPCR实验研究调控机制。结果：我们发现METTL16在PDAC中的表达显著下调，多因素Cox回归分析显示METTL16是PDAC患者的保护性因素。我们还证明了METTL16过表达可抑制PDAC细胞增殖。此外，我们鉴定了METTL16-p21信号轴，METTL16下调导致CDKN1A（p21）抑制。此外，METTL16沉默和过表达实验突显了PDAC中m6A修饰的改变。结论：METTL16通过介导m6A修饰，在p21途径中抑制PDAC细胞增殖，发挥肿瘤抑制的作用。METTL16可能成为PDAC癌变的新型标志物和治疗PDAC的新靶点。