Guan Xiaoqi, Wu Di, Zhu Hongyu, Zhu Biwen, Wang Zhen, Xing Haowei, Zhang Xue, Yan Jiashuai, Guo Yibing, Lu Yuhua
Department of Hepatobiliary and Pancreatic Surgery, Affiliated Hospital of Nantong University, Medical School of Nantong University, 226001 Nantong, Jiangsu, China; Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Medical School of Nantong University, 226001 Nantong, Jiangsu, China.
Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Medical School of Nantong University, 226001 Nantong, Jiangsu, China; Key Laboratory of Neuro-regeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuro-regeneration, Nantong University, 226001 Nantong, Jiangsu, China.
Biomater Adv. 2025 May;170:214215. doi: 10.1016/j.bioadv.2025.214215. Epub 2025 Jan 29.
The distinctive desmoplastic tumor microenvironment (TME) of pancreatic ductal adenocarcinoma (PDAC) is crucial in determining the stemness of tumor cells. And the conventional two-dimensional (2D) culture does not adequately mimic the TME. Therefore, a three-dimensional (3D) PDAC desmoplastic model was constructed using GelMA and HAMA, which provides benefits in terms of simulating both the main components (COL and HA) and the crosslinking of the extracellular matrix. We found that the 3D PDAC desmoplastic model upregulated the expression of the markers for stemness (NANOG and OCT4) and glycolysis (HK2 and GLUT2), and elevated the level of glycolysis, including increased glucose consumption and lactic acid production. Additionally, YAP1 played a crucial role in promoting glycolysis, which boosted stemness. Furthermore, RNA sequencing (RNA-seq) was employed to explore the underlying mechanisms associated with stemness within the 3D desmoplastic model. Subsequent KEGG pathway analysis indicated the activation of the PI3K-AKT signaling pathway, providing insights into the molecular processes at play. Using bioinformatics, qRT-PCR and western blot, we proposed that ITGAV-PI3K-AKT-YAP1 axis may account for the glycolysis mediated the stemness. Collectively, the 3D desmoplastic model may serve as a new platform for understanding the underlying mechanism by which the TME induces stemness.
胰腺导管腺癌（PDAC）独特的促纤维增生性肿瘤微环境（TME）在决定肿瘤细胞的干性方面至关重要。而传统的二维（2D）培养不能充分模拟TME。因此，使用甲基丙烯酰化明胶（GelMA）和透明质酸甲基丙烯酸酯（HAMA）构建了三维（3D）PDAC促纤维增生模型，该模型在模拟主要成分（胶原蛋白和透明质酸）以及细胞外基质交联方面具有优势。我们发现，3D PDAC促纤维增生模型上调了干性标志物（NANOG和OCT4）和糖酵解标志物（己糖激酶2和葡萄糖转运蛋白2）的表达，并提高了糖酵解水平，包括增加葡萄糖消耗和乳酸生成。此外，Yes相关蛋白1（YAP1）在促进糖酵解从而增强干性方面发挥了关键作用。此外，采用RNA测序（RNA-seq）来探索3D促纤维增生模型中与干性相关的潜在机制。随后的京都基因与基因组百科全书（KEGG）通路分析表明PI3K-AKT信号通路被激活，这为其中的分子过程提供了见解。通过生物信息学、定量逆转录聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法，我们提出整合素αV（ITGAV）-PI3K-AKT-YAP1轴可能是糖酵解介导干性的原因。总的来说，3D促纤维增生模型可能成为理解TME诱导干性潜在机制的新平台。
