Augeron C, Laboisse C L
Cancer Res. 1984 Sep;44(9):3961-9.
The human colonic cancer cell line HT29 is undifferentiated in standard culture conditions (Dulbecco's medium:10% fetal bovine serum). These cells were cultured in 5 mM sodium butyrate for 9 days; then they were trypsinized and subcultured in sodium butyrate for an additional 14 days. Multinucleation occurred during this second phase of the treatment. The cells were then transferred to standard medium and multinucleation disappeared. Morphological changes appeared 10 to 12 days after return to standard culture conditions; some cells flattened and became more adherent to the bottom of the flasks. These altered cells divided actively and formed "flat foci" interspersed among the densely packed undifferentiated HT29 cells. This altered phenotype persists after more than 24 months of culture in standard medium. Clonal cell lines were established from these flat foci-forming cells and characterized. These clonal lines exhibited morphological cell polarity defined by an apical cell surface separated by junctional complexes from the basolateral cell surface. Functional differentiation did also occur since some clonal lines formed domes representing active transepithelial transport, and others exhibited massive mucus secretion. In conclusion, our findings indicate that permanently differentiated cell populations emerged in a colonic cancer cell line after sodium butyrate treatment. These new clonal lines will be useful in future models for the study of differentiation programs of both normal and cancerous colonic cells.
人结肠癌细胞系HT29在标准培养条件下（杜尔贝科培养基：10%胎牛血清）未分化。这些细胞在5 mM丁酸钠中培养9天；然后用胰蛋白酶消化，并在丁酸钠中传代培养14天。在处理的第二阶段出现了多核现象。然后将细胞转移到标准培养基中，多核现象消失。回到标准培养条件10至12天后出现形态学变化；一些细胞变平并更紧密地贴附于培养瓶底部。这些改变的细胞积极分裂并形成散布在密集堆积的未分化HT29细胞中的“扁平病灶”。在标准培养基中培养超过24个月后，这种改变的表型仍然存在。从这些形成扁平病灶的细胞建立了克隆细胞系并进行了表征。这些克隆系表现出形态学上的细胞极性，其顶端细胞表面由连接复合体与基底外侧细胞表面分隔开来。功能分化也确实发生了，因为一些克隆系形成了代表活跃跨上皮运输的穹顶，而其他克隆系则表现出大量黏液分泌。总之，我们的研究结果表明，丁酸钠处理后人结肠癌细胞系中出现了永久分化的细胞群体。这些新的克隆系将在未来用于研究正常和癌性结肠细胞分化程序的模型中发挥作用。
