Development of a co-culture system with induced HepG2 cells and K562 cells for examining drug metabolism in vitro. Studies with cyclophosphamide, ondansetron and cisplatin.
We have established a cell co-culture system for assessing potential cytotoxic effects of drugs and their metabolites in vitro. Human hepatoma cells (HepG2) were cultured for 7 days in modified Earle's medium in order to induce their drug metabolising (primarily mixed function oxidase) enzymes. K562 human erythroleukemic cells in Transwells, were used as indicator cells for the cytotoxic effects of cyclophosphamide (CYP) and Ondansetron (OND) and/or their metabolites, produced by induced HepG2 cells in the co-cultures. CYP was found to be approximately 1000 times more toxic to K562 cells when cultured in the presence of induced HepG2 cells. OND, a selective 5-HT3 receptor antagonist which is used as an anti-emetic during chemotherapy, was not found to be cytotoxic in the co-cultures at concentrations as high as 100 microM. Since OND has been particularly useful in relieving vomiting induced by cisplatin (cisPt) chemotherapy, we also examined the effect of cisPt on K562 cells in the presence and absence of OND, and found no evidence that OND significantly enhances the cytotoxic effect of cisPt on these cells alone or in co-cultures with induced HepG2 cells. The induced HepG2 co-culture system uses cells of human origin and clearly has considerable potential for examining the effects of drugs and their metabolites on indicator cells derived from a tissue of choice. This system may be particularly useful in the assessment of metabolism and toxicity of new drugs intended for human use.
Hall
T J
TJ
MD Laboratories, Congleton, Cheshire, U.K.
Cambridge
G
G
James
P R
PR
D000932:Antiemetics / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000008:administration & dosage / Q000378:metabolism* / Q000633:toxicity; D003520:Cyclophosphamide / Q000008:administration & dosage / Q000378:metabolism* / Q000633:toxicity; D004347:Drug Interactions; D006801:Humans; D007093:Imidazoles / Q000008:administration & dosage / Q000378:metabolism* / Q000633:toxicity; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism; D017294:Ondansetron; D014407:Tumor Cells, Cultured / Q000378:metabolism*
D000932:Antiemetics; D007093:Imidazoles; D017294:Ondansetron; D003520:Cyclophosphamide; D002945:Cisplatin