Department of Hepatobiliary and Pancreatic Surgery, Medical School of Nantong University, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu, China.
Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu, China.
ACS Biomater Sci Eng. 2023 Sep 11;9(9):5347-5360. doi: 10.1021/acsbiomaterials.3c00487. Epub 2023 Aug 10.
Pancreatic ductal adenocarcinoma (PDAC) features high recurrence rates and intensified lethality, accompanied by stiffening of the extracellular matrix (ECM) microenvironment, which is mainly due to the deposition, remodeling, and cross-linking of collagen. Boosted stemness plays an essential role during occurrence and progression, which indicates a poor prognosis. Therefore, it is of great importance to understand the effect of the underlying interaction of matrix stiffness and stemness on PDAC. For this purpose, a methacrylated gelatin (GelMA) hydrogel with tunable stiffness was applied for incubating MIA PaCa-2 and PANC-1 cells. The results demonstrated that compared to the soft group (5% GelMA, w/v), the expression of stemness-related genes (SOX2, OCT4, and NANOG) in the stiff group (10% GelMA, w/v) displayed pronounced elevation as well as sphere formation. Intriguingly, we also observed that matrix stiffness regulated autophagy of PDAC, which played a momentous role in stemness promotion. In order to clarify the underlying relationship between matrix stiffness-mediated cell autophagy and stemness, rescue experiments with rapamycin and chloroquine were conducted with transmission electron microscopy, immunofluorescence staining, sphere formation, and qRT-PCR assays to evaluate the level of stemness and autophagy. For exploring the molecular mechanism in depth, RNA-seq and differential expression of miRNAs were carried out, which may sensor and respond to matrix stiffness during the regulation of stemness and autophagy. In conclusion, we validated that blocking autophagy repressed the stemness induced by matrix stiffness in PDAC and provided a potential therapeutic strategy for this aggressive cancer.
胰腺导管腺癌 (PDAC) 的复发率高且致死率强，同时伴有细胞外基质 (ECM) 微环境的硬度增加，这主要是由于胶原蛋白的沉积、重塑和交联。增强的干性在发生和进展中起着至关重要的作用，预示着预后不良。因此，了解基质硬度和干性之间潜在的相互作用对 PDAC 的影响非常重要。为此，应用了一种可调节硬度的甲基丙烯酰化明胶 (GelMA) 水凝胶来孵育 MIA PaCa-2 和 PANC-1 细胞。结果表明，与软组（5% GelMA，w/v）相比，硬组（10% GelMA，w/v）中干性相关基因（SOX2、OCT4 和 NANOG）的表达明显升高，并且形成球体。有趣的是，我们还观察到基质硬度调节 PDAC 的自噬，这在干性促进中起着重要作用。为了阐明基质硬度介导的细胞自噬与干性之间的潜在关系，我们使用雷帕霉素和氯喹进行了挽救实验，通过透射电子显微镜、免疫荧光染色、球体形成和 qRT-PCR 实验评估干性和自噬水平。为了深入探讨分子机制，进行了 RNA-seq 和 miRNA 的差异表达分析，这可能在调节干性和自噬过程中感应和响应基质硬度。总之，我们验证了阻断自噬可以抑制 PDAC 中基质硬度诱导的干性，并为这种侵袭性癌症提供了一种潜在的治疗策略。
