PDF(Pubmed)
Abstract：
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-KrasG12D/+; lox-stop-lox-Trp53R172H/+ 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.
摘要：
胰腺星状细胞(PSC)是癌症相关成纤维细胞(CAF)的来源之一,因此,在胰腺导管腺癌（PDA）中的重要作用。PDA细胞和CAF之间的旁分泌信号已经被广泛研究,然而，对旁分泌串扰的外部影响知之甚少。本研究旨在通过分析PSC基因表达谱，更深入地了解PSC与癌细胞在不同共培养条件下的交流。与p48-Cre肿瘤细胞的两种非接触式共培养模型；lox-stop-lox-KrasG12D/+；lox-stop-lox-Trp53R172H/+小鼠模型（KPC）和鼠PSC通过微孔膜分离，并在不同区室中生长（标准共培养）或在相同膜的不同侧面上生长（反向共培养），已建立。在与KPC细胞共培养后24小时和72小时进行PSCmRNA的RNA测序分析。对于选定的基因，结果通过定量RT-PCR和免疫细胞化学证实。标准共培养在24小时显示19个差异表达基因(DEG)，在72小时显示52°。在反向共培养中，800℃24h和2213℃72h分别富集。PSC在其基因表达谱中表现出极大的异质性；然而，两种共培养的相互调节基因，如VCAN和CHST11，可以识别。VCAN-蛋白质-蛋白质相互作用网络分析揭示了共培养模型之间的几个共享基因，例如SDC4和FN1。总之,根据共培养方法，PSC对癌细胞信号的敏感性不同，随着更接近的转录组变化加剧。
