Regulation of carboxylesterase-2 expression by p53 family proteins and enhanced anti-cancer activities among 5-fluorouracil, irinotecan and doxazolidine prodrug.
OBJECTIVE
For four decades, 5-fluorouracil (5-FU) has been a major anti-cancer medicine. This drug is increasingly used with other anti-cancer agents such as irinotecan. Irinotecan and many others such as PPD (pentyl carbamate of p-aminobenzyl carbamate of doxazolidine) require activation by carboxylesterase-2 (CES2). 5-FU, on the other hand, reportedly induces CES2 in colorectal tumour lines. The aims of this study were to determine the molecular basis for the induction and to ascertain interactive cell-killing activity between 5-FU and ester prodrugs.


METHODS
Colorectal and non-colorectal lines and xenografts were treated with 5-FU and the expression of CES2 was determined. Cell-killing activity of irinotecan and PPD were determined in the presence or absence of CES2 inhibitor. Several molecular experiments were used to determine the molecular basis for the induction.


RESULTS
Without exceptions, robust induction was detected in cell lines expressing functional p53. High-level induction was also detected in xenografts. 5-FU pretreatment significantly increased cell-killing activity of irinotecan and PPD. Molecular experiments established that the induction was achieved by both transactivation and increased mRNA stability through p53. Either p63 or p73, functionally related to p53, did not support the transactivation.


CONCLUSIONS
The results in this study suggest that FOLFIRI, a common regimen combining irinotecan and 5-FU, should switch the dosing sequence, namely from 5-FU to irinotecan, to enhance hydrolytic activation of irinotecan. This modified order likely reduces the dose of anti-cancer agents, thus minimizing overall toxicity. The results also conclude that p53 family members act differently in regulating gene expression.
Department of Biomedical and Pharmaceutical Sciences, Center for Pharmacogenomics and Molecular Therapy, University of Rhode Island, Kingston, RI 02881, USA.
Da
D
Xiao
Dongfang
D
Yang
Liangran
L
Guo
Wei
W
Lu
Margaret
M
Charpentier
Bingfang
B
Yan
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D018938:Caco-2 Cells; D002166:Camptothecin / Q000031:analogs & derivatives* / Q000494:pharmacology; D043182:Carboxylesterase / Q000235:genetics / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases / Q000235:genetics* / Q000378:metabolism*; D045744:Cell Line, Tumor; D004317:Doxorubicin / Q000031:analogs & derivatives* / Q000494:pharmacology; D005472:Fluorouracil / Q000494:pharmacology*; D015972:Gene Expression Regulation, Neoplastic; D019073:HT29 Cells; D056945:Hep G2 Cells; D006801:Humans; D000077146:Irinotecan; D051379:Mice; D010080:Oxazoles / Q000494:pharmacology*; D020871:RNA Stability; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D023041:Xenograft Model Antitumor Assays
D000970:Antineoplastic Agents; D010080:Oxazoles; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; C545998:doxazolidine; D000077146:Irinotecan; D004317:Doxorubicin; D002265:Carboxylic Ester Hydrolases; C517204:CES2 protein, human; D043182:Carboxylesterase; C500145:Ces2c protein, mouse; D005472:Fluorouracil; D002166:Camptothecin