title: Histone H3 messenger RNA in situ hybridization correlates with in vivo bromodeoxyuridine labeling of S-phase cells in rat colonic epithelium.
abstract: Measurements of cell cycle phase fractions, particularly S-phase, are useful for studies of cell biology and carcinogenesis. Up-regulation of histone gene expression is tightly coupled to the G1-S-phase transition of the cell cycle, and mRNA levels rise 30-100-fold during S-phase. Labeling of histone H3 mRNA using in situ hybridization (ISH) was assessed as a measure of S-phase cells and compared with that found using in vivo 5-bromodeoxyuridine (BrdUrd) labeling in formalin-fixed rat colonic crypts under baseline, modified 72-h starvation, and 24-h refeeding conditions. The labeling index scored in single-labeled sections by histone H3 ISH tightly correlated with that found by in vivo BrdUrd labeling (r = 0.99, p < 0.0001) and clearly discriminated between the control, starved, and refed states (P < 0.001). In 180 crypt sections double labeled using histone H3 ISH and BrdUrd, 92% of 1572 labeled cells exhibited both nuclear BrdUrd and cytoplasmic histone H3 label. It is concluded that histone H3 ISH is an accurate measure of the S-phase fraction and provides an alternative to in vivo BrdUrd labeling in rat colon. This finding warrants validation in human studies.
authors.affiliation: Department of Gastrointestinal Medical Oncology and Digestive Diseases, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
authors.forename: H
authors.initials: H
authors.lastname: Konishi
authors.forename: G
authors.initials: G
authors.lastname: Steinbach
authors.forename: N H
authors.initials: NH
authors.lastname: Terry
authors.forename: J J
authors.initials: JJ
authors.lastname: Lee
authors.forename: J A
authors.initials: JA
authors.lastname: Dubin
authors.forename: G A
authors.initials: GA
authors.lastname: Glober
authors.forename: K
authors.initials: K
authors.lastname: Fujita
authors.forename: D
authors.initials: D
authors.lastname: Spaulding
authors.forename: L
authors.initials: L
authors.lastname: Cass
authors.forename: W N
authors.initials: WN
authors.lastname: Hittelman
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D001973:Bromodeoxyuridine / Q000378:metabolism*; D002455:Cell Division / Q000502:physiology; D003106:Colon / Q000166:cytology* / Q000378:metabolism; D003110:Colonic Neoplasms / Q000378:metabolism / Q000473:pathology; D004195:Disease Models, Animal; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D006657:Histones / Q000032:analysis* / Q000235:genetics; D017403:In Situ Hybridization; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism; D008297:Male; D051379:Mice; D011230:Precancerous Conditions / Q000378:metabolism / Q000473:pathology; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D011916:Rats, Inbred F344; D015203:Reproducibility of Results; D016196:S Phase / Q000502:physiology*; D013217:Starvation / Q000378:metabolism
chemical_list: D015415:Biomarkers; D006657:Histones; D012333:RNA, Messenger; D001973:Bromodeoxyuridine