title: Hydrophobic bile acid-induced micronuclei formation, mitotic perturbations, and decreases in spindle checkpoint proteins: relevance to genomic instability in colon carcinogenesis.
abstract: We show, for the first time, that hydrophobic bile acids cause aberrations of the mitotic machinery of colon cells that can give rise to aneuploidy, the chromosomal perturbations common in colon tumors. First, we show that DOC induces a statistically significant fourfold increase in the number of micronuclei in NCM-460 cells (a noncancerous colon cell line) and a threefold increase in the number of micronuclei in binucleated HT-29 colon cancer cells using the cytokinesis block micronucleus assay. Second, we observed mitotic aberrations after DOC treatment, including improper alignment of chromosomes at the metaphase plate, lagging chromosomes during anaphase, anaphase/telophase chromatin bridges, multipolar divisions, and formation of polynucleated cells. It was determined that there was a statistically significant threefold increase in the number of aberrant metaphases after short-term and long-term exposure of HT-29 and HCT-116 cells, respectively. Third, we showed with Western blots and immunohistochemistry that a likely basis for these mitosis-related perturbations included decreased expression of the spindle checkpoint proteins, Mad2, BubR1, and securin. Fourth, results of DOC treatment on nocodazole-challenged cells further indicated deficiencies in activation of the spindle assembly checkpoint. This study provides mechanisms by which hydrophobic bile acids can induce genomic instability in colon epithelial cells.
authors.affiliation: Department of Cell Biology and Anatomy, College of Medicine, University of Arizona, Tucson, AZ 85724-5044, USA. cpayne@email.arizona.edu
authors.forename: Claire M
authors.initials: CM
authors.lastname: Payne
authors.forename: Cheray
authors.initials: C
authors.lastname: Crowley-Skillicorn
authors.forename: Carol
authors.initials: C
authors.lastname: Bernstein
authors.forename: Hana
authors.initials: H
authors.lastname: Holubec
authors.forename: Mary Pat
authors.initials: MP
authors.lastname: Moyer
authors.forename: Harris
authors.initials: H
authors.lastname: Bernstein
mesh_terms: D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000209:etiology* / Q000235:genetics; D003840:Deoxycholic Acid / Q000633:toxicity*; D042822:Genomic Instability / Q000187:drug effects*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D048629:Micronuclei, Chromosome-Defective; D008938:Mitosis / Q000187:drug effects*; D015739:Nocodazole / Q000494:pharmacology; D008941:Spindle Apparatus / Q000187:drug effects*
chemical_list: D003840:Deoxycholic Acid; D015739:Nocodazole