Bile acid-induced colon cancer cell proliferation

Summary

Principal Investigator: Jean Pierre Raufman
Affiliation: University of Maryland
Country: USA
Abstract: The central hypothesis to be tested in this proposal is that bile acids induce colon cancer cell proliferation by interaction with M3 muscarinic receptors (M3R), thereby causing transactivation of epidermal growth factor receptors (EGFR). To elucidate post-receptor signaling that results in bile acid-induced colon cancer cell proliferation and to determine the requirement for, and mechanism of, transactivation of EGFR the following Specific Aims are proposed: 1. To elucidate further post-receptor signal transduction pathways which mediate cholinergic agonist-induced transactivation of EGFR and stimulate colon cancer cell proliferation. 1a. For use in the proposed studies, in addition to cells that naturally express M3R and EGFR, colon cancer cells will be transfected with cDNA clones for M3R and/or EGFR. 1b. The mechanism of cholinergic agonist-induced cell proliferation will be studied using immunoblotting to probe for activated proteins in the p44/42 MARK, p38 MARK and JNK pathways, and by examining the roles of phospholipase C and protein kinase C activation and Ca2+ mobilization.1c. The requirement for cholinergic agonist-induced transactivation of EGFR will be confirmed by examining EGFR phosphorylation and by using EGFR inhibitors, antisense oligonucleotides, and dominant negative mutants.2. To determine the requirement for co-expression of M3R and EGFR and delineate the molecular mechanisms whereby bile acids regulate colon cancer cell proliferation. 2a. Bile acid-induced post-receptor signaling and the requirement for co-expression of M3R and EGFR will be determined using radioligand binding assays, immunoblotting for activated p44/42 MARK, p38 MARK and JNK cascade proteins, and by using EGFR inhibitors, antisense oligonucleotides, and dominant negative mutants. 2b. Bile acid-induced activation and expression of transcription factors (p90RSK, p38 MARK and JNK) and genes (CREB, NF-kappaB, c-Fos and c-Jun) related to colon cancer proliferation will be elucidated. 3. To determine the molecular mechanism in human colon cancer cells of bile acid-induced transactivation of EGFR. 3a) Expression and release of EGFR ligands, including HB-EGF, will be determined using immunoblots, northern blots, and RT-PCR, and the dependence of bile acid-induced EGFR transactivation on release of EGFR ligands will be determined using EGFR antibodies, metalloproteinase inhibitors, and specific antibodies and inhibitors for EGFR. 3b) Colon cancer cell metalloproteases will be identified by immunoblotting and in situ hybridization and the role of these enzymes in mediating bile acid-induced HB-EGF release will be determined by using metalloprotease inhibitors and antisera, and by knocking down metalloprotease expression. 3c) The mechanism whereby bile acids activate metalloproteases will be determined by exploring the roles of PKC, Ca2+, and Src in bile acid-induced HB-EGF release.
Funding Period: 2005-06-28 - 2010-05-31
more information: NIH RePORT

Top Publications

  1. pmc Akt-dependent NF-kappaB activation is required for bile acids to rescue colon cancer cells from stress-induced apoptosis
    Jasleen Shant
    Division of Gastroenterology and Hepatology, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA
    Exp Cell Res 315:432-50. 2009
  2. pmc Divergent effects of muscarinic receptor subtype gene ablation on murine colon tumorigenesis reveals association of M3R and zinc finger protein 277 expression in colon neoplasia
    Kunrong Cheng
    Department of Medicine, Division of Gastroenterology and Hepatology and Program in Oncology, Marlene and Stewart Greenebaum Cancer Center, VA Maryland Health Care System and University of Maryland School of Medicine, Baltimore, MD 21201 1595, USA
    Mol Cancer 13:77. 2014
  3. pmc The Cdc42/Rac nucleotide exchange factor protein β1Pix (Pak-interacting exchange factor) modulates β-catenin transcriptional activity in colon cancer cells: evidence for direct interaction of β1PIX with β-catenin
    Ahmed Chahdi
    From the Division of Gastroenterology and Hepatology, Department of Medicine
    J Biol Chem 288:34019-29. 2013
  4. pmc Effects of modulating M3 muscarinic receptor activity on azoxymethane-induced liver injury in mice
    Sandeep Khurana
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System and University of Maryland School of Medicine, Baltimore, MD, United States
    Biochem Pharmacol 86:329-38. 2013
  5. pmc Cholinergic muscarinic receptor activation augments murine intestinal epithelial cell proliferation and tumorigenesis
    Zhongsheng Peng
    Division of Gastroenterology and Hepatology, Veterans Affairs Maryland Health Care System, University of Maryland School of Medicine, Baltimore, MD, USA
    BMC Cancer 13:204. 2013
  6. pmc Src-mediated cross-talk between farnesoid X and epidermal growth factor receptors inhibits human intestinal cell proliferation and tumorigenesis
    Zhongsheng Peng
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System, University of Maryland School of Medicine, Baltimore, United States of America
    PLoS ONE 7:e48461. 2012
  7. pmc Bile acids regulate cardiovascular function
    Sandeep Khurana
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System and University of Maryland School of Medicine, Baltimore, MD, USA
    Clin Transl Sci 4:210-8. 2011
  8. pmc Muscarinic receptor agonists stimulate matrix metalloproteinase 1-dependent invasion of human colon cancer cells
    Jean Pierre Raufman
    Division of Gastroenterology and Hepatology, University of Maryland School of Medicine, Baltimore, MD 21201, USA
    Biochem Biophys Res Commun 415:319-24. 2011
  9. pmc Effects of deoxycholylglycine, a conjugated secondary bile acid, on myogenic tone and agonist-induced contraction in rat resistance arteries
    Sandeep Khurana
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System and University of Maryland School of Medicine, Baltimore, Maryland, United States of America
    PLoS ONE 7:e32006. 2012
  10. pmc Src-mediated aryl hydrocarbon and epidermal growth factor receptor cross talk stimulates colon cancer cell proliferation
    Guofeng Xie
    Division of Gastroenterology and Hepatology, Univ of Maryland School of Medicine, 22 S Greene St, N3W62, Baltimore, MD 21201, USA
    Am J Physiol Gastrointest Liver Physiol 302:G1006-15. 2012

Scientific Experts

  • Jean Pierre Raufman
  • Guofeng Xie
  • Sandeep Khurana
  • Kunrong Cheng
  • Zhongsheng Peng
  • Jasleen Shant
  • Ahmed Chahdi
  • Nirish Shah
  • Jonathon Heath
  • Angelica Belo
  • Cinthia Drachenberg
  • James P Hamilton
  • Cinthia B Drachenberg
  • Jennifer Timmons
  • Bernard S Marasa
  • Jian Ying Wang
  • Roxana Samimi
  • Richard J Davis
  • George Nomikos
  • Mark Wade
  • Dale A Chatfield
  • Christine A Packard
  • Lee R Hagey
  • Alan F Hofmann
  • Terrance L Griffin
  • Susan Hogan
  • Joseph H Steinbach

Detail Information

Publications21

  1. pmc Akt-dependent NF-kappaB activation is required for bile acids to rescue colon cancer cells from stress-induced apoptosis
    Jasleen Shant
    Division of Gastroenterology and Hepatology, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA
    Exp Cell Res 315:432-50. 2009
    ..These findings provide a mechanism whereby bile acids increase resistance of colon cancer to chemotherapy and radiation...
  2. pmc Divergent effects of muscarinic receptor subtype gene ablation on murine colon tumorigenesis reveals association of M3R and zinc finger protein 277 expression in colon neoplasia
    Kunrong Cheng
    Department of Medicine, Division of Gastroenterology and Hepatology and Program in Oncology, Marlene and Stewart Greenebaum Cancer Center, VA Maryland Health Care System and University of Maryland School of Medicine, Baltimore, MD 21201 1595, USA
    Mol Cancer 13:77. 2014
    ..In mice, ablating Chrm3, the gene encoding M3R, robustly attenuates intestinal tumor formation. Here we investigated the effects of Chrm1 gene ablation, alone and in combination with Chrm3 ablation...
  3. pmc The Cdc42/Rac nucleotide exchange factor protein β1Pix (Pak-interacting exchange factor) modulates β-catenin transcriptional activity in colon cancer cells: evidence for direct interaction of β1PIX with β-catenin
    Ahmed Chahdi
    From the Division of Gastroenterology and Hepatology, Department of Medicine
    J Biol Chem 288:34019-29. 2013
    ..Altogether, these findings suggest that β1Pix functions as a transcriptional regulator of β-catenin signaling through direct interaction with β-catenin, an action that may be functionally relevant to colon cancer biology. ..
  4. pmc Effects of modulating M3 muscarinic receptor activity on azoxymethane-induced liver injury in mice
    Sandeep Khurana
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System and University of Maryland School of Medicine, Baltimore, MD, United States
    Biochem Pharmacol 86:329-38. 2013
    ..These novel observations provide a proof-of-principle that selectively stimulating M3R activation to prevent or diminish liver injury is a therapeutic strategy worthy of further investigation. ..
  5. pmc Cholinergic muscarinic receptor activation augments murine intestinal epithelial cell proliferation and tumorigenesis
    Zhongsheng Peng
    Division of Gastroenterology and Hepatology, Veterans Affairs Maryland Health Care System, University of Maryland School of Medicine, Baltimore, MD, USA
    BMC Cancer 13:204. 2013
    ..Previously, we showed that M3 muscarinic receptor (M3R; gene name Chrm3) deficiency attenuates murine intestinal neoplasia, supporting the hypothesis that muscarinic receptors play an important role in intestinal tumorigenesis...
  6. pmc Src-mediated cross-talk between farnesoid X and epidermal growth factor receptors inhibits human intestinal cell proliferation and tumorigenesis
    Zhongsheng Peng
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System, University of Maryland School of Medicine, Baltimore, United States of America
    PLoS ONE 7:e48461. 2012
    ..Collectively these data support the novel conclusion that in human colon cancer cells Src-mediated cross-talk between FXR and EGFR modulates ERK phosphorylation, thereby regulating intestinal cell proliferation and tumorigenesis...
  7. pmc Bile acids regulate cardiovascular function
    Sandeep Khurana
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System and University of Maryland School of Medicine, Baltimore, MD, USA
    Clin Transl Sci 4:210-8. 2011
    ..This review focuses on BAs as newly recognized signaling molecules that modulate cardiovascular function...
  8. pmc Muscarinic receptor agonists stimulate matrix metalloproteinase 1-dependent invasion of human colon cancer cells
    Jean Pierre Raufman
    Division of Gastroenterology and Hepatology, University of Maryland School of Medicine, Baltimore, MD 21201, USA
    Biochem Biophys Res Commun 415:319-24. 2011
    ....
  9. pmc Effects of deoxycholylglycine, a conjugated secondary bile acid, on myogenic tone and agonist-induced contraction in rat resistance arteries
    Sandeep Khurana
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System and University of Maryland School of Medicine, Baltimore, Maryland, United States of America
    PLoS ONE 7:e32006. 2012
    ..Although in both health and disease serum glycine-conjugated BAs are more abundant than taurine-conjugated BAs, their effects on myogenic tone (MT), a key determinant of systemic vascular resistance (SVR), have not been examined...
  10. pmc Src-mediated aryl hydrocarbon and epidermal growth factor receptor cross talk stimulates colon cancer cell proliferation
    Guofeng Xie
    Division of Gastroenterology and Hepatology, Univ of Maryland School of Medicine, 22 S Greene St, N3W62, Baltimore, MD 21201, USA
    Am J Physiol Gastrointest Liver Physiol 302:G1006-15. 2012
    ..These data support novel observations that, in human colon cancer cells, Src-mediated cross talk between aryl hydrocarbon and EGFR results in ERK1/2 activation, thereby stimulating cell proliferation...
  11. pmc Muscarinic receptor subtype-3 gene ablation and scopolamine butylbromide treatment attenuate small intestinal neoplasia in Apcmin/+ mice
    Jean Pierre Raufman
    Department of Medicine, Division of Gastroenterology and Hepatology, VA Maryland Health Care System and University of Maryland School of Medicine, Baltimore, MD 21201, USA
    Carcinogenesis 32:1396-402. 2011
    ....
  12. pmc Muscarinic receptor agonists stimulate human colon cancer cell migration and invasion
    Angelica Belo
    Division of Gastroenterology and Hepatology, Univ of Maryland School of Medicine, Baltimore, 21201, USA
    Am J Physiol Gastrointest Liver Physiol 300:G749-60. 2011
    ..In concert with previous observations, these findings indicate that muscarinic receptor signaling plays a key role in colon cancer cell proliferation, survival, migration, and invasion...
  13. pmc Scopolamine treatment and muscarinic receptor subtype-3 gene ablation augment azoxymethane-induced murine liver injury
    Sandeep Khurana
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System and University of Maryland School of Medicine, Baltimore, Maryland 21201, USA
    J Pharmacol Exp Ther 333:639-49. 2010
    ..We conclude that Chrm3 plays a critical role in the liver injury response by modulating hepatocyte proliferation and apoptosis...
  14. pmc Matrix metalloproteinase-7-catalyzed release of HB-EGF mediates deoxycholyltaurine-induced proliferation of a human colon cancer cell line
    Kunrong Cheng
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System and Program in Oncology, Greenebaum Cancer Center, University of Maryland School of Medicine, 22 S Greene Street, N3W62, Baltimore, MD 21201, USA
    Biochem Pharmacol 73:1001-12. 2007
    ..These findings provide strong evidence that in H508 human colon cancer cells, DCT-induced transactivation of EGFR is mediated by MMP-7-catalyzed release of the EGFR ligand HB-EGF...
  15. ncbi Human cecal bile acids: concentration and spectrum
    James P Hamilton
    Division of Gastroenterology and Hepatology, Department of Medicine, Veterans Affairs Maryland Health Care System and University of Maryland School of Medicine, 22 S Greene Street, Baltimore, MD 21201, USA
    Am J Physiol Gastrointest Liver Physiol 293:G256-63. 2007
    ..In the human cecum, deconjugation and dehydroxylation of bile acids are nearly complete, resulting in most bile acids being in unconjugated form at submicellar and subsecretory concentrations...
  16. pmc Deoxycholyltaurine rescues human colon cancer cells from apoptosis by activating EGFR-dependent PI3K/Akt signaling
    Jean Pierre Raufman
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System and Program in Oncology, Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, Maryland
    J Cell Physiol 215:538-49. 2008
    ..Collectively, these results indicate that DCT-induced activation of post-EGFR PI3K/Akt signaling stimulates both colon cancer cell survival and proliferation...
  17. pmc Genetic ablation of M3 muscarinic receptors attenuates murine colon epithelial cell proliferation and neoplasia
    Jean Pierre Raufman
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System and Program in Oncology, Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, USA
    Cancer Res 68:3573-8. 2008
    ..001). Compared with WT, azoxymethane-treated M(3)R-deficient mice have attenuated epithelial cell proliferation, tumor number, and size. M(3)R and post-M(3)R signaling are novel therapeutic targets for colon cancer...
  18. pmc Acetylcholine release by human colon cancer cells mediates autocrine stimulation of cell proliferation
    Kunrong Cheng
    Division of Gastroenterology and Hepatology, Veterans Affairs Maryland Healthcare System, University of Maryland School of Medicine, Department of Pathology, 22 S Greene St, N3W62, Baltimore, MD 21201, USA
    Am J Physiol Gastrointest Liver Physiol 295:G591-7. 2008
    ..005). We conclude that ACh is an autocrine growth factor in colon cancer. Mechanisms that regulate colon epithelial cell production and release of ACh warrant further investigation...
  19. pmc Muscarinic receptors and ligands in cancer
    Nirish Shah
    Division of Gastroenterology and Hepatology, Univ of Maryland School of Medicine, 22 South Greene St, N3W62, Baltimore, MD 21201, USA
    Am J Physiol Cell Physiol 296:C221-32. 2009
    ..Dissecting cellular mechanisms necessary for muscarinic receptor activation as well as those needed for acetylcholine production and release will identify multiple novel targets for cancer therapy...
  20. pmc Acetylcholine-induced activation of M3 muscarinic receptors stimulates robust matrix metalloproteinase gene expression in human colon cancer cells
    Guofeng Xie
    Division of Gastroenterology and Hepatology, Veterans Administration Maryland Health Care System and Program in Oncology, Greenbaum Cancer Center, University of Maryland School of Medicine, 22 S Greene St, N3W62, Baltimore, MD 21201, USA
    Am J Physiol Gastrointest Liver Physiol 296:G755-63. 2009
    ..ACh-induced activation of EGFR and downstream ERK signaling also regulates transcriptional activation of MMP7, thereby identifying a novel feed-forward mechanism for neoplastic cell proliferation...
  21. ncbi Bile acid-induced proliferation of a human colon cancer cell line is mediated by transactivation of epidermal growth factor receptors
    Kunrong Cheng
    Division of Gastroenterology and Hepatology, VA Maryland Health Care System and Program in Oncology, Greenebaum Cancer Center, University of Maryland School of Medicine, 22 S Green Street, N3W62 Baltimore, MD 21201, USA
    Biochem Pharmacol 70:1035-47. 2005
    ..Collectively, these results suggest that in this colon cancer cell line, bile acid-induced colon cancer cell proliferation is M3R-dependent and is mediated by transactivation of EGFR...