Department of Gastrointestinal Surgery, The Affiliated Huaian No. 1 People's Hospital of Nanjing Medical University, Huai'an, Jiangsu, 223300, People's Republic of China.
Department of Vascular, Huaian Hospital Affiliated to Xuzhou Medical University, Huai'an, Jiangsu, 223300, People's Republic of China.
Sci Rep. 2024 Mar 19;14(1):6568. doi: 10.1038/s41598-024-57099-7.
While Phorbol-12-myristate-13-acetate-induced protein 1 (Noxa/PMAIP1) assumes a pivotal role in numerous tumors, its clinical implications and underlying mechanisms of gastric cancer (GC) are yet enigmatic. In this investigation, our primary objective was to scrutinize the clinical relevance and potential mechanisms of Noxa in gastric cancer. Immunohistochemical analysis was conducted on tissue microarrays comprising samples from a meticulously characterized cohort of 84 gastric cancer patients, accompanied by follow-up data, to assess the expression of Noxa. Additionally, Noxa expression levels in gastric cancer clinical samples and cell lines were measured through quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis. The effect of Noxa expression on the prognosis of patients with gastric cancer was evaluated using Kaplan-Meier survival. Further insight into the role of Noxa in driving gastric cancer progression was gained through an array of experimental techniques, including cell viability assays (CCK8), plate cloning assays, transwell assays, scratch assays, and real-time cell analysis (RTCA). Potential upstream microRNAs (miRNAs) that might modulate Noxa were identified through rigorous bioinformatics analysis, substantiated by luciferase reporter assays and Western blot experiments. Additionally, we utilized RNA sequencing, qRT-PCR, and Western blot to identify proteins binding to Noxa and potential downstream target. Finally, we utilized BALB/c nude mice to explore the role of Noxa in vivo. Our investigation unveiled a marked downregulation of Noxa expression in gastric cancer and underscored its significance as a pivotal prognostic factor influencing overall survival (OS). Noxa overexpression exerted a substantial inhibitory effect on the proliferation, migration and invasion of GC cells. Bioinformatic analysis and dual luciferase reporter assays unveiled the capacity of hsa-miR-200b-3p to interact with the 3'-UTR of Noxa mRNA, thereby orchestrating a downregulation of Noxa expression in vitro, consequently promoting tumor progression in GC. Our transcriptome analysis, coupled with mechanistic validation, elucidated a role for Noxa in modulating the expression of ZNF519 in the Mitophagy-animal pathway. The depletion of ZNF519 effectively reversed the oncogenic attributes induced by Noxa. Upregulation of Noxa expression suppressed the tumorigenesis of GC in vivo. The current investigation sheds light on the pivotal role of the hsa-miR-200b-3p/Noxa/ZNF519 axis in elucidating the pathogenesis of gastric cancer, offering a promising avenue for targeted therapeutic interventions in the management of this challenging malignancy.
虽然佛波醇-12-肉豆蔻酸-13-醋酸酯诱导蛋白 1(Noxa/PMAIP1)在许多肿瘤中起着关键作用,但它在胃癌(GC)中的临床意义和潜在机制仍然是个谜。在这项研究中,我们的主要目的是研究 Noxa 在胃癌中的临床相关性和潜在机制。我们对包含 84 名胃癌患者的精心表征队列的组织微阵列进行了免疫组织化学分析,并进行了随访数据,以评估 Noxa 的表达。此外,我们通过定量实时聚合酶链反应(qRT-PCR)和 Western blot 分析测量了胃癌临床样本和细胞系中 Noxa 的表达水平。通过 Kaplan-Meier 生存分析评估 Noxa 表达对胃癌患者预后的影响。通过一系列实验技术,包括细胞活力测定(CCK8)、平板克隆测定、Transwell 测定、划痕测定和实时细胞分析(RTCA),进一步深入了解 Noxa 在推动胃癌进展中的作用。通过严格的生物信息学分析鉴定了可能调节 Noxa 的潜在上游 microRNAs(miRNAs),并通过荧光素酶报告基因测定和 Western blot 实验进行了验证。此外,我们还利用 RNA 测序、qRT-PCR 和 Western blot 鉴定了与 Noxa 结合的蛋白和潜在的下游靶标。最后,我们利用 BALB/c 裸鼠在体内探索了 Noxa 的作用。我们的研究揭示了 Noxa 在胃癌中的表达明显下调,并强调了其作为影响总生存(OS)的关键预后因素的重要性。Noxa 过表达对 GC 细胞的增殖、迁移和侵袭具有显著的抑制作用。生物信息学分析和双荧光素酶报告基因测定揭示了 hsa-miR-200b-3p 与 Noxa mRNA 的 3'-UTR 相互作用的能力,从而在体外下调 Noxa 的表达,进而促进 GC 中的肿瘤进展。我们的转录组分析,结合机制验证,阐明了 Noxa 在调节 Mitophagy-animal 通路中 ZNF519 表达中的作用。ZNF519 的耗尽有效地逆转了 Noxa 诱导的致癌特性。Noxa 表达的上调抑制了体内 GC 的肿瘤发生。目前的研究揭示了 hsa-miR-200b-3p/Noxa/ZNF519 轴在阐明胃癌发病机制中的关键作用,为该挑战性恶性肿瘤的靶向治疗干预提供了有希望的途径。