Department of Visceral, Vascular and Endocrine Surgery, University Hospital Halle, Martin-Luther-University Halle-Wittenberg, Halle, Germany.
Biocenter, Martin-Luther-University Halle-Wittenberg, Halle, Germany.
Sci Rep. 2024 Aug 4;14(1):18030. doi: 10.1038/s41598-024-68148-6.
Pancreatic stellate cells (PSC) are one source of cancer-associated fibroblasts (CAF) and play, therefore, an essential role in pancreatic ductal adenocarcinoma (PDA). Paracrine signalling between PDA cells and CAF has been widely studied, yet external influences on paracrine crosstalk are poorly understood. This study aimed to gain a deeper insight into the communication of PSC and cancer cells under different co-culture conditions via analysis of PSC gene expression profiles. Two contactless co-culture models with tumor cells from the p48-Cre; lox-stop-lox-Kras; lox-stop-lox-Trp53 mouse model (KPC) and murine PSC separated through a microporous membrane and grown in different compartments (standard co-culture) or on different sides of the same membrane (inverse co-culture), were established. RNA-Sequencing analysis of PSC mRNA was performed 24 h and 72 h after co-culture with KPC cells. For selected genes, results were confirmed by quantitative RT-PCR and immunocytochemistry. Standard co-culture displayed 19 differentially expressed genes (DEG) at 24 h and 52 DEG at 72 h. In inverse co-culture, 800 DEG at 24 h and 2213 DEG at 72 h were enriched. PSC showed great heterogeneity in their gene expression profiles; however, mutually regulated genes of both co-cultures, such as VCAN and CHST11, could be identified. VCAN-protein-protein interaction-network analysis revealed several shared genes between co-culture models, such as SDC4 and FN1. In conclusion, PSC show a varying susceptibility to cancer cell signals depending on the co-culture method, with intensified transcriptome changes with closer proximity.
胰腺星状细胞（PSC）是癌相关成纤维细胞（CAF）的一个来源，因此在胰腺导管腺癌（PDA）中起着至关重要的作用。已经广泛研究了 PDA 细胞和 CAF 之间的旁分泌信号，但对外源性旁分泌串扰的影响知之甚少。本研究旨在通过分析 PSC 的基因表达谱，更深入地了解不同共培养条件下 PSC 和癌细胞之间的通讯。通过微孔膜将肿瘤细胞与胰腺星状细胞分离，并在不同隔室（标准共培养）或同一膜的不同侧（反向共培养）中生长，建立了两种无接触共培养模型，其中一种是 p48-Cre; lox-stop-lox-Kras; lox-stop-lox-Trp53 小鼠模型（KPC）的肿瘤细胞，另一种是鼠胰腺星状细胞。在与 KPC 细胞共培养 24 小时和 72 小时后，对 PSC 的 mRNA 进行 RNA-Sequencing 分析。对于选定的基因，通过定量 RT-PCR 和免疫细胞化学法对结果进行了验证。标准共培养在 24 小时和 72 小时时分别有 19 个差异表达基因（DEG）和 52 个 DEG。在反向共培养中，在 24 小时时有 800 个 DEG，在 72 小时时有 2213 个 DEG 被富集。PSC 的基因表达谱具有很大的异质性；然而，两种共培养物中相互调节的基因，如 VCAN 和 CHST11，可以被识别。VCAN 蛋白-蛋白相互作用网络分析显示了两种共培养模型之间的一些共享基因，如 SDC4 和 FN1。总之，PSC 对癌细胞信号的敏感性取决于共培养方式，随着距离的接近，转录组的变化会加剧。
