Huang Yishan, Yang Ting, Yang Chen, Tang Bo, Su Bo, Yang Xiaojun
Engineering Research Center of Key Technique for Biotherapy of Guangdong Province, Shantou University Medical College, Shantou, 515041, China.
Department of Hepatobiliary Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, China.
Sci Rep. 2025 May 22;15(1):17870. doi: 10.1038/s41598-025-02586-8.
Pancreatic cancer remains one of the leading causes of mortality worldwide, largely due to the limitations of current clinical strategies for its treatment. As a result, identifying genetic alterations and potential therapeutic targets could offer new opportunities for improving the diagnosis and treatment of pancreatic cancer. The identification of differentially expressed genes (DEGs) and subsequent analyses, including signaling pathway enrichment, functional classification, and protein-protein interaction (PPI) network construction, were conducted using three public datasets: GSE32676, GSE71989, and GSE16515. Kaplan-Meier survival curves and receiver operating characteristic (ROC) curves were employed to investigate the correlation between hub genes and clinicopathological features in pancreatic cancer patients. Genetic alterations were analyzed using the CBioPortal web tool. Cell proliferation was assessed through CCK-8, colony formation, and EdU assays. Tumor migration, invasion, and angiogenesis were evaluated using transwell and tube formation assays, respectively. Protein and mRNA expression levels were measured via western blot analysis and qPCR assays. The subcutaneous xenografted nude mice models were generated to evaluate the potential effect of miR-485-3p/MELK cascade on tumor growth in vivo. Our analysis revealed that MELK expression is positively correlated with poor prognosis in patients with pancreatic cancer. The overexpression or knockdown of MELK significantly influences cell proliferation, tumor metastasis, and angiogenesis across various pancreatic cancer cell lines. Furthermore, we identified that miR-485-3p regulates MELK expression by directly targeting the MELK 3'UTR binding site in pancreatic cancer cells, which subsequently impacts tumor progression. Additionally, our findings demonstrate that the miR-485-3p/MELK cascade is closely associated with tumor progression in pancreatic cancer cells. Mechanistically, the miR-485-3p/MELK cascade promotes the phosphorylation of Akt to regulate pancreatic cancer cell progression, metastasis, and angiogenesis. Furthermore, overexpression of miR-485-3p inhibits the tumor growth induced by MELK overexpression in subcutaneous xenograft model. MiR-485-3p/MELK cascade may serve as a promising biomarker and therapeutic target for the diagnosis and treatment of pancreatic cancer.
胰腺癌仍然是全球主要的死亡原因之一，这主要归因于当前胰腺癌临床治疗策略的局限性。因此，识别基因改变和潜在的治疗靶点可为改善胰腺癌的诊断和治疗提供新的机会。使用三个公共数据集GSE32676、GSE71989和GSE16515进行差异表达基因（DEG）的识别以及后续分析，包括信号通路富集、功能分类和蛋白质-蛋白质相互作用（PPI）网络构建。采用Kaplan-Meier生存曲线和受试者工作特征（ROC）曲线来研究胰腺癌患者中核心基因与临床病理特征之间的相关性。使用CBioPortal网络工具分析基因改变。通过CCK-8、集落形成和EdU试验评估细胞增殖。分别使用Transwell试验和管形成试验评估肿瘤迁移、侵袭和血管生成。通过蛋白质印迹分析和qPCR试验测量蛋白质和mRNA表达水平。建立皮下异种移植裸鼠模型以评估miR-485-3p/MELK级联对体内肿瘤生长的潜在影响。我们的分析表明，MELK表达与胰腺癌患者的不良预后呈正相关。MELK的过表达或敲低显著影响各种胰腺癌细胞系中的细胞增殖、肿瘤转移和血管生成。此外，我们发现miR-485-3p通过直接靶向胰腺癌细胞中的MELK 3'UTR结合位点来调节MELK表达，进而影响肿瘤进展。此外，我们的研究结果表明，miR-485-3p/MELK级联与胰腺癌细胞中的肿瘤进展密切相关。从机制上讲，miR-485-3p/MELK级联促进Akt的磷酸化以调节胰腺癌细胞的进展、转移和血管生成。此外，在皮下异种移植模型中，miR-485-3p的过表达抑制了MELK过表达诱导的肿瘤生长。MiR-485-3p/MELK级联可能是胰腺癌诊断和治疗的一个有前景的生物标志物和治疗靶点。
