Transport and metabolism of polyamines in wild and multidrug resistant human leukemia (K 562) cells.
Multidrug resistance (MDR) can be defined as the resistance of cancer cells not just to chemotherapeutic agents to which they have been exposed but also to other apparently unrelated compounds. This MDR phenotype is commonly associated with the high expression of levels of 170 kDa P-glycoprotein, encoded by MDR genes. In the present study, the uptake kinetics of polyamines and their biosynthesis were studied in wild and multidrug resistant (MDR) K 562 cells in culture. The rate (Vmax) of polyamine uptake was significantly lower in MDR cells than that in wild type cells, whereas the Km for the uptake was not significantly different in these cells, suggesting that polyamine transporter is not modified in MDR cells, though their different physiological state influences the uptake process. In a 32 h chase, the transported radioactive polyamines were gradually interconverted. [14C]putrescine was converted into [14C]spermidine following between 15 min and 32 h of culture, and into [14C]-spermine after 16 h of culture, in both the cell types; however, the levels of interconverted radioactive polyamines were always lower in MDR cells as compared with wild type cells. Similarly, internalized [14C]spermidine was converted into [14C]spermine, but not into [14C]putrescine in both the cells types. [14C]spermidine is metabolized into [14C]spermine after 4 h of culture in wild type cells, whereas in MDR cells the interconversion of [14C]spermidine into [14C]spermine is seen only after 16 h of culture. Blocking of the transmembrane drug efflux pump, expressed in the MDR cells, by preincubation in the presence of verapamil, did not influence the uptake of either of the two polyamines (putrescine and spermidine) by MDR cells. On the contrary, this kind of preincubation of wild type cells in the presence of verapamil significantly increased the uptake of these two polyamines. The levels of intracellular polyamine contents in MDR cells were always lower than those in the parental cell line. These results demonstrate that MDR cells are defective in both the uptake of polyamines and their biosynthesis as compared with wild type cells.
Khan
N A
NA
Laboratoire de Biologie Cellulaire, Centre Hospitalier Universitaire, Faculté de Médecine, Rennes, France.
Fardel
O
O
Havouis
R
R
Fauchet
R
R
Moulinoux
J P
JP
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D001692:Biological Transport / Q000187:drug effects; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002455:Cell Division; D004351:Drug Resistance; D015870:Gene Expression; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism* / Q000473:pathology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D011700:Putrescine / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D013095:Spermidine / Q000378:metabolism*; D013997:Time Factors; D014407:Tumor Cells, Cultured; D014700:Verapamil / Q000494:pharmacology
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D014700:Verapamil; D013095:Spermidine; D011700:Putrescine