Organization of the endoplasmic reticulum-Golgi system is related to the state of enterocytic differentiation of human HT-29 cells

Abstract

The organization of the endoplasmic reticulum (ER)-intermediate compartment (IC)-Golgi system was studied in tumoral HT-29 cells. Depending on the culture conditions, these cells are either undifferentiated or exhibit enterocytic differentiation after reaching confluence. In differentiated HT-29 cells, these organelles were organized as in most cell types. They displayed a classical structure and appeared associated with microtubules, as nocodazole altered both their structure and intracellular localization. Likewise, membrane dynamics of the Golgi appeared normal: as in many other cells, brefeldin A (BFA) induced retrograde transport from the Golgi to the ER, demonstrated by tubulation of the Golgi elements and shift of the galactosyltransferase activity from the Golgi- to the RER-enriched fraction, isolated by subcellular fractionation. In contrast, atypical features were observed in undifferentiated HT-29 cells: the Golgi structure exhibited abnormal swellings; the IC elements were very rare. Only cytochalasin D altered the structure and intracellular localization of the three organelles, suggesting that they were associated with microfilaments instead of microtubules. The membrane dynamics were unusual: brefeldin A led to a vesiculation of the Golgi elements with a slowed-down retrograde transport of galactosyltransferase. HT-29 cells engaged in the differentiation process, but which were still undifferentiated, showed mainly the features of undifferentiated cells, with a few characteristics of differentiated cells. These results indicate that the structure of the Golgi apparatus, IC and ER, their relationships to cytoskeletal elements and membrane dynamics depend on the state of differentiation of HT-29 cells. Although they are tumoral, differentiated HT-29 cells exhibit features observed in non-tumoral polarized epithelial cells. On the contrary, undifferentiated HT-29 cells display important abnormalities that may be related to their metastatic properties.