Studies on Epithelial-mesenchymal Interaction in Gastrointestinal Tract.

在这项研究中，我们使用三维培养系统研究了胃肠道(GI)上皮细胞和成纤维细胞之间的上皮-间充质相互作用。所有被检测的成纤维细胞系(NIH 3T3、Sweiss 3T3、MRC-5、NRK)都能促进未转化的Gl管上皮细胞(RGM-1、IEC-6和原代胎兔胃上皮细胞)的生长/存活。未转化的胃上皮细胞株和原代胃上皮细胞分泌一种促进成纤维细胞生长/存活和形态转化的活性，而转化的上皮细胞系(MKN-28、45、DLD-1、HepG2、COS-1)则没有这种活性。未转化的胃上皮细胞系RGM-1产生一种促进成纤维细胞和Ito细胞生长/存活和形态转化的活性(S)，并刺激PC-12神经细胞突起生长。为了纯化这种活性(S)，我们从RGM-1细胞的大规模培养中收集了30份L的无血清条件培养液。活性通过肝素亲和、离子交换、凝胶过滤和反相色谱结合FPLC系统进行纯化。纯化后的活性分子量为30 kDa，其周边区氨基酸序列为新的。

In this study, we investigated epithelial-mesenchymal interaction between gastrointestinal (GI) epithelial cells and fibroblasts using a three dimensional culture system. All the fibroblast cell line examined (NIH 3T3, Sweiss 3T3, MRC-5, NRK) produced an activity that promote growth/survival of non-transformed epithelial cells from the Gl tract (RGM-1, IEC-6, and primary fetal rabbit gastic epithelial cells). Non-trasformed epithelial cell lines and primary gastric epithelial cells secreted an activity that promote growth/survival and morphological transformation of the fibroblasts, whereas this activity was lost in transformed epithelial cell lines (MKN-28,45, DLD-1, HepG2, COS-1).RGM-1, a non-transformed gastric epithelial cell line, produced an activity (s) that promote growth/survival and morphological transformation of fibroblasts and Ito cell and stimulates neurite outgrowth of PC-12 neuronal cell. To purify this activity (s), we collected 30 L of serum-free conditioned medium from large scale culture of RGM-1 cell. The activity was purified by a combination of heparin affinity, ion exchange, gel filtration, and reverse phase chromatography using a FPLC system. The purified activity has a molecular weight of 30 kDa and its pertial amino acid sequence was novel.