Glucuronidation as a mechanism of intrinsic drug resistance in human colon cancer: reversal of resistance by food additives.
Colon cancer exhibits inherent insensitivity to chemotherapy by mechanisms that are poorly characterized. We have shown that human colon cancer cells are efficient in drug conjugation catalyzed by UDP-glucuronosyltransferases (UGTs) and now report on the role of glucuronidation in de novo resistance to two topoisomerase I inhibitors. Identification of the UGT responsible for glucuronidation of SN-38 and the anthraquinone NU/ICRF 505 was achieved by first using a panel of human cDNA-expressed isozymes to measure conjugating activity. HT29 colon cancer cells were then probed by reverse transcriptase-PCR, Western Blot analysis, and liquid chromatography with mass spectrometry for their profile and activity of UGT isozymes and screened for effective inhibitors of glucuronidation. Expression analysis was also conducted in colon cancer biopsies and paired adjacent normal colon specimens. UGT1A9 was identified as the isozyme catalyzing biotransformation of the two compounds in HT29 cells and propofol as an effective competitive inhibitor of this metabolism. Inhibition of glucuronidation resulted in up to a 5-fold enhancement in drug activity. The majority of colon cancer biopsies studies expressed UGT protein at levels greater than in HT29 cells but with marked interpatient variations and proficiently glucuronidated the two anticancer drugs. A range of UGT aglycones were capable of modulating glucuronidation in the biopies with octylgallate being 10-fold more potent (ID(50) 24 microM) than propofol. In a subset of tumors (33%), UGT protein levels and activity exceeded that of paired normal colon. Glucuronidation may represent a mechanism of intrinsic drug resistance in colon cancer open to modulation by a range of food additives and proprietary medicines.
Cummings
Jeffrey
J
Cancer Research UK, Edinburgh Oncology Unit, Western General Hospital, Edinburgh, United Kingdom. jcummings@picr.man.ac.uk
Ethell
Brian T
BT
Jardine
Lesley
L
Boyd
Gary
G
Macpherson
Janet S
JS
Burchell
Brian
B
Smyth
John F
JF
Jodrell
Duncan I
DI
D000880:Anthraquinones / Q000378:metabolism*; D001706:Biopsy; D002166:Camptothecin / Q000031:analogs & derivatives* / Q000378:metabolism*; D002384:Catalysis; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000378:metabolism*; D019008:Drug Resistance, Neoplasm / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000378:metabolism; D005503:Food Additives / Q000494:pharmacology*; D020719:Glucuronides / Q000378:metabolism*; D014453:Glucuronosyltransferase / Q000378:metabolism; D006801:Humans; D000077146:Irinotecan; D007527:Isoenzymes / Q000378:metabolism; D015742:Propofol / Q000378:metabolism / Q000494:pharmacology; D059004:Topoisomerase I Inhibitors; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism*
C441475:7-ethyl-10-hydroxycamptothecin glucuronide; D000880:Anthraquinones; D004791:Enzyme Inhibitors; D005503:Food Additives; D020719:Glucuronides; D007527:Isoenzymes; C100645:NU-ICRF 505; D059004:Topoisomerase I Inhibitors; D014443:Tyrosine; D000077146:Irinotecan; D014453:Glucuronosyltransferase; D002166:Camptothecin; D015742:Propofol