Abstract：
Cancer stem cells (CSC) are thought to be responsible for cancer phenotypes and cellular heterogeneity. Here we demonstrate that the human colon cancer cell line DLD1 contains two types of CSC-like cells that undergo distinct morphogenesis in the reconstituted basement membrane gel Matrigel. In our method with cancer cell spheroids, the parent cell line (DLD1-P) developed grape-like budding structures, whereas the other (DLD1-Wm) and its single-cell clones dynamically developed worm-like ones. Gene expression analysis suggested that the former mimicked intestinal crypt-villus morphogenesis, while the latter mimicked embryonic hindgut development. The organoids of DLD1-Wm cells rapidly extended in two opposite directions by expressing dipolar proteolytic activity. The invasive morphogenesis required the expression of MMP-2 and CD133 genes and ROCK activity. These cells also exhibited gastrula-like morphogenesis even in two-dimensional cultures without Matrigel. Moreover, the two DLD1 cell lines showed clear differences in cellular growth, tumor growth and susceptibility to paclitaxel. This study also provides a simple organoid culture method for human cancer cell lines. HT-29 and other cancer cell lines underwent characteristic morphogenesis in direct contact with normal fibroblasts. Such organoid cultures would be useful for investigating the nature of CSCs and for screening anti-cancer drugs. Our results lead to the hypothesis that CSC-like cells with both invasive activity and a fetal phenotype, i.e,. oncofetal CSCs, are generated in some types of colon cancers.
摘要：
癌症干细胞(CSC)被认为是癌症表型和细胞异质性的原因。在这里，我们证明了人结肠癌细胞系DLD1包含两种类型的CSC样细胞，它们在重构的基底膜凝胶Matrigel中经历不同的形态发生。在我们用癌细胞球体的方法中，亲本细胞系（DLD1-P）发育出葡萄样的出芽结构，而另一个（DLD1-Wm）及其单细胞克隆动态地发展了蠕虫样克隆。基因表达分析表明，前者模仿肠道隐窝绒毛形态发生，而后者模仿胚胎后肠发育。DLD1-Wm细胞的类器官通过表达偶极蛋白水解活性在两个相反的方向上快速延伸。侵袭性形态发生需要MMP-2和CD133基因的表达和ROCK活性。即使在没有Matrigel的二维培养物中，这些细胞也表现出胃肠病样形态发生。此外，两种DLD1细胞系在细胞生长方面表现出明显的差异，肿瘤生长和对紫杉醇的敏感性。这项研究还为人类癌细胞系提供了一种简单的类器官培养方法。HT-29和其他癌细胞系经历了与正常成纤维细胞直接接触的特征性形态发生。此类类器官培养物可用于研究CSC的性质和用于筛选抗癌药物。我们的结果导致假设CSC样细胞具有侵袭活性和胎儿表型，即，.癌胎儿CSC，在某些类型的结肠癌中产生。
