Suramin-induced differentiation of the human colic adenocarcinoma cell clone HT29-D4 in serum-free medium.
The clonal cell line HT29-D4 was able to grow in a completely defined medium containing EGF, selenous acid, and transferrin in the presence of the anti-helminthic drug suramin. In the absence of suramin, the kinetics of cell growth and the cell density obtained were dependent on the external EGF concentration. In the presence of suramin, cell density reached a plateau independent of EGF concentration above 50 ng/ml. At the morphological level, suramin allowed hemicyst formation in the cell monolayer. The cells were polarized with a well-ordered brush border facing the culture medium and mature junctional complexes that divided the cell membrane in two distinct domains. The carcinoembryonic antigen was found to be restricted to the apical membrane domain while the major histocompatibility molecules HLA-ABC were segregated within the basolateral domain. The electrical parameters of suramin-treated cells grown on permeable filters were measured and demonstrated that the cell monolayer was electrically active. These properties were never found in the absence of the drug. Moreover, the vasoactive intestinal polypeptide (VIP) was able to induce a dramatic increase in cAMP only when it was added, in agreement with the localization of the VIP receptor, in the lower compartment of the culture chamber. In conclusion we described for the first time conditions allowing the growth of functionally differentiated human colic cell monolayers in chemically defined medium. This model will contribute to a better understanding of suramin action and of the mechanisms involved in cell polarization.
Fantini
J
J
Centre National de la Recherche Scientifique, Institut de Chimie Biologique, Faculté St. Charles, France.
Verrier
B
B
Robert
C
C
Pic
P
P
Pichon
J
J
Mauchamp
J
J
Marvaldi
J
J
D000230:Adenocarcinoma / Q000473:pathology*; D002272:Carcinoembryonic Antigen / Q000032:analysis; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects; D002999:Clone Cells; D003110:Colonic Neoplasms / Q000473:pathology*; D003470:Culture Media; D000242:Cyclic AMP / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000494:pharmacology; D006680:HLA Antigens / Q000032:analysis; D006801:Humans; D013498:Suramin / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D014660:Vasoactive Intestinal Peptide / Q000494:pharmacology
D002272:Carcinoembryonic Antigen; D003470:Culture Media; D006680:HLA Antigens; D014660:Vasoactive Intestinal Peptide; D013498:Suramin; D004815:Epidermal Growth Factor; D000242:Cyclic AMP