在结直肠癌（CRC）中，肿瘤旁的正常组织（NAT）与肿瘤之间有着积极的相互作用。大肠中的成年干细胞在结肠上皮的发育中起着至关重要的作用。然而，在肿瘤微环境中，从NAT派生的结肠干细胞发生了哪些变化尚不清楚。本研究利用肠道干细胞培养系统，分别培养了来自NAT、与CRC组织成对的结肠细胞，以及来自健康组织（HLT）的细胞。通过比较NAT、HLT和CRC组织的克隆的集落形成能力和分化能力，来比较克隆的功能。利用这些克隆的RNA高通量测序来鉴定NAT派生的克隆的分子特征。利用来自HLT和CRC的克隆的共培养来评估分子变化。我们发现，NAT派生的克隆的形态特征、集落形成能力和分化能力与HLT派生的克隆一致。然而，NAT派生的克隆在分子水平上发生了变化。许多基因在NAT中特异性激活。NAT派生的克隆丰富了与炎症和纤维化相关的通路，包括上皮间质转变（EMT）通路和TGF-beta信号通路。我们的结果还证实，NAT派生的克隆能够在小鼠中招募成纤维细胞。此外，当与肿瘤细胞共培养时，HLT派生的克隆表达FOSB高。我们的结果表明，肿瘤微环境中来自NAT的结肠干细胞在分子水平上发生了变化，并且这些分子特征可以在体外维持，从而诱导纤维化和炎症反应。视频摘要。©2023. 作者。

In colorectal cancer (CRC), the normal tissue adjacent to tumor (NAT) communicates actively with the tumor. Adult stem cells from the colon play a crucial role in the development of the colonic epithelium. In the tumor microenvironment, however, it is unclear what changes have occurred in colonic stem cells derived from NAT.Using an intestinal stem cell culture system, we cultured colonic cells from NAT and paired CRC tissue, as well as cells from healthy tissue (HLT). Clonogenicity and differentiation ability were used to compare the function of clones from NAT, HLT and CRC tissues. RNA high-throughput sequencing of these clones was used to identify the molecular characteristics of NAT-derived clones. Coculture of clones from HLT and CRC was used to assess molecular changes.We found that the morphological characteristics, clonogenic ability, and differentiation ability of NAT-derived clones were consistent with those of HLT-derived clones. However, NAT-derived clones changed at the molecular level. A number of genes were specifically activated in NAT. NAT-derived clones enriched pathways related to inflammation and fibrosis, including epithelial mesenchymal transition (EMT) pathway and TGF-beta signaling pathway. Our results also confirmed that NAT-derived clones could recruit fibroblasts in mice. In addition, HLT-derived clones showed high expression of FOSB when cocultured with tumor cells.Our results demonstrate that colonic stem cells from NAT in the tumor microenvironment undergo changes at the molecular level, and these molecular characteristics can be maintained in vitro, which can induce fibrosis and an inflammatory response. Video Abstract.© 2023. The Author(s).