Bossard C, Jarry A, Colombeix C, Bach-Ngohou K, Moreau A, Loussouarn D, Mosnier J-F, Laboisse C L
INSERM, U539, Nantes, France.
J Clin Pathol. 2006 Jul;59(7):706-10. doi: 10.1136/jcp.2005.030452. Epub 2006 Feb 3.
Microscopic evaluation of mitotic figures is a routine procedure in the assessment of the histoprognostic grade of tumours. Nevertheless, their count may be fraught with difficulties. As histone H3 phosphorylation at serine 10 is closely linked to chromosomal condensation, a new monoclonal antibody directed to phosphorylated histone H3 (PPH3) was recently proposed to detect mitotic cells.
To test the reliability of this antibody in detecting and counting mitotic figures in sections of breast adenocarcinomas, because of the importance of mitotic count in histoprognostic grading.
The pattern of PPH3 staining in formalin-fixed paraffin wax-embedded tissues, including normal tissues and a series of 39 breast adenocarcinomas, was examined. A new computer-assisted method was also developed for determining the mitotic index.
In all tissues tested, PPH3-labelled mitotic figures were easily detected, allowing a rapid identification of the area of highest mitotic activity. In breast carcinomas, a strong correlation was observed between PPH3-stained and haematoxylin and eosin-stained mitotic counts (r = 0.86, p<0.0001). Counting of prophase nuclei that coexpress cyclin B1, a marker of the G2/M phase, was possible by PPH3 staining; its accuracy led us to reconsider the tumour grade in three cases. Finally, an automatic computer-assisted method was designed for assessing mitotic index with confocal microscopy and image-analysis software.
有丝分裂图像的显微镜评估是肿瘤组织预后分级评估中的常规程序。然而，对其进行计数可能存在困难。由于组蛋白H3丝氨酸10位点的磷酸化与染色体浓缩密切相关，最近有人提出一种针对磷酸化组蛋白H3（PPH3）的新型单克隆抗体来检测有丝分裂细胞。
鉴于有丝分裂计数在组织预后分级中的重要性，测试该抗体在检测和计数乳腺腺癌切片中有丝分裂图像方面的可靠性。
检查了PPH3在福尔马林固定石蜡包埋组织（包括正常组织和39例乳腺腺癌系列组织）中的染色模式。还开发了一种新的计算机辅助方法来确定有丝分裂指数。
在所有测试组织中，PPH3标记的有丝分裂图像易于检测，从而能够快速识别有丝分裂活性最高的区域。在乳腺癌中，观察到PPH3染色的有丝分裂计数与苏木精和伊红染色的有丝分裂计数之间存在强相关性（r = 0.86，p<0.0001）。通过PPH3染色可以对同时表达细胞周期蛋白B1（G2/M期标志物）的前期细胞核进行计数；其准确性使我们重新考虑了3例病例的肿瘤分级。最后，设计了一种自动计算机辅助方法，用于通过共聚焦显微镜和图像分析软件评估有丝分裂指数。
