title: Accuracy of histone H3 messenger RNA in situ hybridization for the assessment of cell proliferation in human tissues.
abstract: Histone H3 mRNA in situ hybridization was compared to a reference method, iododeoxyuridine (IdUrd) immunohistochemistry of tissues labeled in vivo, as a means for assessing the proportion of S-phase cells (labeling index, LI) in oral tumor and normal mucosa. Paraffin sections from 16 patients with oral squamous cell carcinoma were studied. Patients received an IdUrd infusion before the biopsy was taken. Tissue sections were coded before counting the percentages of S-phase cells. A high correlation was found between the results obtained by the two techniques. The average histone H3 and IdUrd LIs of the tumors were 28.5 +/- 2.4% and 29.2 +/- 2.7%, respectively (P = 0.85), with a Spearman correlation coefficient r = 0.95 (P < 0. 0001). The histone H3 LI of the basal layer of normal mucosa was 3.1 +/- 0.8%, whereas the IdUrd LI was 2.7 +/- 0.9% (P = 0.74), with r = 0.78 (P = 0.004). In the suprabasal layers, these parameters were 21. 3 +/- 2.3% and 23.9 +/- 3.2%, respectively (P = 0.56), with r = 0.93 (P < 0.0001). In sections stained for both histone H3 and IdUrd, most cells were double labeled, with very few cells containing only one of the labels. In some specimens, large areas of H3-stained cells did not contain IdUrd-labeled cells, suggesting that during the IdUrd infusion, the precursor did not reach these areas. Two specimens were histone H3 negative. They were also negative when hybridized with beta-actin probe, indicating degradation of mRNAs in these samples. The results of this study demonstrate that the histone H3 mRNA in situ hybridization performed in human formalin-fixed, paraffin-embedded tissues provides the same data as does labeling the tumors in vivo with halogenated pyrimidine.
authors.affiliation: Rush Cancer Institute and Pathology Department, Rush-Presbyterian-St. Lukes Medical Center, Chicago, Illinois 60612, USA.
authors.forename: V
authors.initials: V
authors.lastname: Kotelnikov
authors.forename: L
authors.initials: L
authors.lastname: Cass
authors.forename: J S
authors.initials: JS
authors.lastname: Coon
authors.forename: D
authors.initials: D
authors.lastname: Spaulding
authors.forename: H D
authors.initials: HD
authors.lastname: Preisler
mesh_terms: D001706:Biopsy; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000473:pathology*; D002455:Cell Division; D006258:Head and Neck Neoplasms / Q000235:genetics / Q000473:pathology; D006657:Histones / Q000235:genetics*; D006801:Humans; D007065:Idoxuridine; D017403:In Situ Hybridization / Q000379:methods; D008940:Mitotic Index; D009061:Mouth Mucosa / Q000378:metabolism / Q000473:pathology*; D009062:Mouth Neoplasms / Q000235:genetics* / Q000473:pathology*; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D015203:Reproducibility of Results; D016196:S Phase
chemical_list: D006657:Histones; D012333:RNA, Messenger; D007065:Idoxuridine