Kotelnikov V, Cass L, Coon J S, Spaulding D, Preisler H D
Rush Cancer Institute and Pathology Department, Rush-Presbyterian-St. Lukes Medical Center, Chicago, Illinois 60612, USA.
Clin Cancer Res. 1997 May;3(5):669-73.
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.
将组蛋白H3 mRNA原位杂交与一种参考方法进行比较，该参考方法是对体内标记组织进行碘脱氧尿苷（IdUrd）免疫组化，以此作为评估口腔肿瘤和正常黏膜中S期细胞比例（标记指数，LI）的一种手段。研究了16例口腔鳞状细胞癌患者的石蜡切片。患者在活检前接受IdUrd输注。在计算S期细胞百分比之前，对组织切片进行编码。发现两种技术获得的结果之间具有高度相关性。肿瘤的组蛋白H3和IdUrd平均LI分别为28.5±2.4%和29.2±2.7%（P = 0.85），Spearman相关系数r = 0.95（P < 0.0001）。正常黏膜基底层的组蛋白H3 LI为3.1±0.8%，而IdUrd LI为2.7±0.9%（P = 0.74），r = 0.78（P = 0.004）。在基底上层，这些参数分别为21.3±2.3%和23.9±3.2%（P = 0.56），r = 0.93（P < 0.0001）。在同时用组蛋白H3和IdUrd染色的切片中，大多数细胞被双重标记，只有极少数细胞仅含有一种标记。在一些标本中，大片H3染色的细胞不含IdUrd标记的细胞，这表明在IdUrd输注期间，前体未到达这些区域。两个标本组蛋白H3呈阴性。当与β-肌动蛋白探针杂交时，它们也呈阴性，表明这些样本中的mRNA发生了降解。本研究结果表明，在人福尔马林固定、石蜡包埋组织中进行的组蛋白H3 mRNA原位杂交所提供的数据与用卤代嘧啶对肿瘤进行体内标记所提供的数据相同。
