Phosphodiesterase in human colon carcinoma cell line CaCo-2 in culture.
In a series of in vitro experiments we characterised the relationship between DNA distribution in the G1, S and G2/M phases of cell cycle and PDE and GST activity in CaCo-2 cells. The DNA distribution in CaCo-2 cells, was assessed by flow cytometry, with fluorescent dyes at different time points of culture. The exponential increase in cell number continued until day 10 when there was cell saturation. The effect of medium replacement on PDE activity was assayed in the first 10 h after medium replacement. The 6th hour is the time at which PDE activity was found to be highest. We have assayed the PDE enzyme with cGMP and cAMP as substrates. Only cAMP was consumed from this enzyme. We found a very close correlation between the DNA distribution in the various phases of the cell cycle and the PDE activity. PDE activity was very high during the active replication phase, whereas GST activity was high after confluency.
Savini
F
F
Chair of Clinical Pathology, University G. D'Annunzio, Chieti, Italy.
Berardi
S
S
Tatone
D
D
Spoto
G
G
D002277:Carcinoma / Q000201:enzymology* / Q000473:pathology; D002453:Cell Cycle; D002454:Cell Differentiation; D002455:Cell Division; D003110:Colonic Neoplasms / Q000201:enzymology* / Q000473:pathology; D004273:DNA, Neoplasm / Q000096:biosynthesis; D005434:Flow Cytometry; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D013379:Substrate Specificity; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D005982:Glutathione Transferase; D010727:Phosphoric Diester Hydrolases