Histone H3 messenger RNA in situ hybridization correlates with in vivo bromodeoxyuridine labeling of S-phase cells in rat colonic epithelium.
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.
Konishi
H
H
Department of Gastrointestinal Medical Oncology and Digestive Diseases, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Steinbach
G
G
Terry
N H
NH
Lee
J J
JJ
Dubin
J A
JA
Glober
G A
GA
Fujita
K
K
Spaulding
D
D
Cass
L
L
Hittelman
W N
WN
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
D015415:Biomarkers; D006657:Histones; D012333:RNA, Messenger; D001973:Bromodeoxyuridine