Carboxylesterases Expressed in Human Colon Tumor Tissue and Their Role in CPT-11 Hydrolysis

Sonal P. Sanghani, Sara K. Quinney, Tyler B. Fredenburg, Zejin Sun, Wilhelmina I. Davis, Daryl J. Murry, Oscar W. Cummings, David E. Seitz, William F. Bosron

Research output: Contribution to journalArticle

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Abstract

Purpose: The purpose is to develop new analytical methods to study the expression profile of CPT-11 carboxylesterases and topoisomerase I in colon tumor samples and understand the impact of their expression on CPT-11 metabolism in chemotherapy. Experimental Design: We investigated 24 colon tumors for expression of carboxylesterases CES1A1, CES2, CES3, hBr-3, and topoisomerase I genes by real-time PCR and correlated the gene expression with activity assays. The relative abundance of the carboxylesterase isoenzymes and topoisomerase I genes was determined by real-time PCR. Activity assays performed on colon tumor extracts included CPT-11 hydrolase, 4-methylumbelliferyl acetate hydrolase, and topoisomerase I activity assays. Additionally, nondenaturing activity gel electrophoresis with activity staining showed the distribution of carboxylesterases. Results: We detect the expression of CES1A1, CES2, and CES3 carboxylesterase genes in human colon tumors. We were unable to detect the hBr-3 (also called hCE-3) in human liver, colon, or brain. We find large interindividual variation, ≥150-fold, for both CES1A1 and CES3 genes, 23-fold for CES2, and 66-fold for topoisomerase I. Only CES2 gene expression correlated with the carboxylesterase activity assays (P < 0.01) with CPT-11 and 4-methylumbelliferyl acetate as substrates. Nondenaturing activity gel electrophoresis showed that CES2 was the most predominant activity. Topoisomerase I gene expression significantly correlated with topoisomerase I activity (P < 0.01) in the colon tumors, but interindividual variation was very high. Conclusions: We conclude that CES2 is the most abundant carboxylesterase in colon tumors that is responsible for CPT-11 hydrolysis. This pilot study reinforces the hypothesis that there is a large interindividual variation in expression of carboxylesterases that may contribute to variation in therapeutic outcome and/or toxicity of CPT-11 therapy for colon cancer.

Original languageEnglish (US)
Pages (from-to)4983-4991
Number of pages9
JournalClinical Cancer Research
Volume9
Issue number13
StatePublished - Oct 15 2003

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irinotecan
Carboxylic Ester Hydrolases
Type I DNA Topoisomerase
Colon
Hydrolysis
Carboxylesterase
Neoplasms
Hydrolases
Gene Expression
Genes
Electrophoresis
Real-Time Polymerase Chain Reaction
Gels
Colonic Neoplasms
Isoenzymes

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Sanghani, S. P., Quinney, S. K., Fredenburg, T. B., Sun, Z., Davis, W. I., Murry, D. J., ... Bosron, W. F. (2003). Carboxylesterases Expressed in Human Colon Tumor Tissue and Their Role in CPT-11 Hydrolysis. Clinical Cancer Research, 9(13), 4983-4991.

Carboxylesterases Expressed in Human Colon Tumor Tissue and Their Role in CPT-11 Hydrolysis. / Sanghani, Sonal P.; Quinney, Sara K.; Fredenburg, Tyler B.; Sun, Zejin; Davis, Wilhelmina I.; Murry, Daryl J.; Cummings, Oscar W.; Seitz, David E.; Bosron, William F.

In: Clinical Cancer Research, Vol. 9, No. 13, 15.10.2003, p. 4983-4991.

Research output: Contribution to journalArticle

Sanghani, SP, Quinney, SK, Fredenburg, TB, Sun, Z, Davis, WI, Murry, DJ, Cummings, OW, Seitz, DE & Bosron, WF 2003, 'Carboxylesterases Expressed in Human Colon Tumor Tissue and Their Role in CPT-11 Hydrolysis', Clinical Cancer Research, vol. 9, no. 13, pp. 4983-4991.
Sanghani SP, Quinney SK, Fredenburg TB, Sun Z, Davis WI, Murry DJ et al. Carboxylesterases Expressed in Human Colon Tumor Tissue and Their Role in CPT-11 Hydrolysis. Clinical Cancer Research. 2003 Oct 15;9(13):4983-4991.
Sanghani, Sonal P. ; Quinney, Sara K. ; Fredenburg, Tyler B. ; Sun, Zejin ; Davis, Wilhelmina I. ; Murry, Daryl J. ; Cummings, Oscar W. ; Seitz, David E. ; Bosron, William F. / Carboxylesterases Expressed in Human Colon Tumor Tissue and Their Role in CPT-11 Hydrolysis. In: Clinical Cancer Research. 2003 ; Vol. 9, No. 13. pp. 4983-4991.
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T1 - Carboxylesterases Expressed in Human Colon Tumor Tissue and Their Role in CPT-11 Hydrolysis

AU - Sanghani, Sonal P.

AU - Quinney, Sara K.

AU - Fredenburg, Tyler B.

AU - Sun, Zejin

AU - Davis, Wilhelmina I.

AU - Murry, Daryl J.

AU - Cummings, Oscar W.

AU - Seitz, David E.

AU - Bosron, William F.

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N2 - Purpose: The purpose is to develop new analytical methods to study the expression profile of CPT-11 carboxylesterases and topoisomerase I in colon tumor samples and understand the impact of their expression on CPT-11 metabolism in chemotherapy. Experimental Design: We investigated 24 colon tumors for expression of carboxylesterases CES1A1, CES2, CES3, hBr-3, and topoisomerase I genes by real-time PCR and correlated the gene expression with activity assays. The relative abundance of the carboxylesterase isoenzymes and topoisomerase I genes was determined by real-time PCR. Activity assays performed on colon tumor extracts included CPT-11 hydrolase, 4-methylumbelliferyl acetate hydrolase, and topoisomerase I activity assays. Additionally, nondenaturing activity gel electrophoresis with activity staining showed the distribution of carboxylesterases. Results: We detect the expression of CES1A1, CES2, and CES3 carboxylesterase genes in human colon tumors. We were unable to detect the hBr-3 (also called hCE-3) in human liver, colon, or brain. We find large interindividual variation, ≥150-fold, for both CES1A1 and CES3 genes, 23-fold for CES2, and 66-fold for topoisomerase I. Only CES2 gene expression correlated with the carboxylesterase activity assays (P < 0.01) with CPT-11 and 4-methylumbelliferyl acetate as substrates. Nondenaturing activity gel electrophoresis showed that CES2 was the most predominant activity. Topoisomerase I gene expression significantly correlated with topoisomerase I activity (P < 0.01) in the colon tumors, but interindividual variation was very high. Conclusions: We conclude that CES2 is the most abundant carboxylesterase in colon tumors that is responsible for CPT-11 hydrolysis. This pilot study reinforces the hypothesis that there is a large interindividual variation in expression of carboxylesterases that may contribute to variation in therapeutic outcome and/or toxicity of CPT-11 therapy for colon cancer.

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