McClelland R A, Finlay P, Walker K J, Nicholson D, Robertson J F, Blamey R W, Nicholson R I
Tenovus Institute for Cancer Research, Department of Surgery, City Hospital, Nottingham, United Kingdom.
Cancer Res. 1990 Jun 15;50(12):3545-50.
Frozen sections of breast tumor tissue have been stained using an immunoperoxidase [estrogen receptor (ER)-immunocytochemistry] kit incorporating a monoclonal antiserum [H222] to visualize nuclear human ERs. Quantitation of specific staining has been performed by manual procedures using optical microscopy and by a computer-assisted image analysis system (CAS 100). Initial investigations with a test panel of ER-immunocytochemistry-positive tumors revealed a good qualitative agreement between CAS and manual assessments. Reduced variance was, however, observed between quantified ER-immunocytochemistry results from four experienced investigators using the CAS analysis. An extended study confirmed the relationships between CAS and manual methods of assessment. These findings were evident when studies were scored either by assessment of the percentage of positively stained cells (n = 92; r = 0.919; P less than 0.01) or by H-score calculations (n = 92; r = 0.913; P less than 0.01). A good correlation was also found between CAS quantification and the results of an ER enzyme immunoassay of 48 primary breast cancer specimens (r = 0.715; P less than 0.05). In 49 cases it was possible to relate CAS-defined ER status and levels to the subsequent response of patients to endocrine therapy. ER was assessed on specimens obtained prior to commencement of treatments for recurrent breast cancer. Presuming the presence of ER to be a prerequisite for successful therapy, very good correlations between response and both status and levels of positivity were recorded. None of 16 patients with CAS-ER-negative tumors responded to treatment, while 16 of 33 (48.4%) CAS-ER-positive patients achieved an objective response according to International Union Against Cancer criteria. A relationship between response and the degree of CAS-ER positivity was obtained when the CAS score divisions of 0, 1-100, and greater than 100 (response rates, 0, 41, and 64%, respectively) were used. These data demonstrate that automated image analysis offers a reliable, reproducible procedure for quantifying ER in immunocytochemically stained sections. It has potential advantages over manual procedures, providing less opportunity for subjective influences in scoring sections. Future advances in software design should further reduce elements of subjectivity and increase both the speed and reliability of results. We anticipate image analysis becoming a valuable tool in investigations concerning, for example, the influence of heterogeneity of steroid receptor distribution on the rate of recurrence of breast cancer after mastectomy and in the clinical course of the disease.
乳腺肿瘤组织的冰冻切片已使用包含抗人雌激素受体单克隆抗血清[H222]的免疫过氧化物酶（雌激素受体免疫细胞化学）试剂盒进行染色，以显示细胞核内的人雌激素受体。通过光学显微镜的手工操作以及计算机辅助图像分析系统（CAS 100）对特异性染色进行定量。对一组雌激素受体免疫细胞化学阳性肿瘤的初步研究表明，CAS分析与手工评估之间在定性方面具有良好的一致性。然而，在使用CAS分析对四位经验丰富的研究人员的雌激素受体免疫细胞化学定量结果之间观察到方差减小。一项扩展研究证实了CAS与手工评估方法之间的关系。当通过评估阳性染色细胞百分比（n = 92；r = 0.919；P < 0.01）或H评分计算（n = 92；r = 0.913；P < 0.01）对研究进行评分时，这些结果很明显。在对48例原发性乳腺癌标本进行的雌激素受体酶免疫测定结果与CAS定量之间也发现了良好的相关性（r = 0.715；P < 0.05）。在49例病例中，可以将CAS定义的雌激素受体状态和水平与患者随后对内分泌治疗的反应联系起来。在复发性乳腺癌治疗开始前获取的标本上评估雌激素受体。假定雌激素受体的存在是成功治疗的先决条件，记录到反应与阳性状态和水平之间具有非常好的相关性。16例CAS雌激素受体阴性肿瘤患者均未对治疗产生反应，而33例（48.4%）CAS雌激素受体阳性患者中有16例根据国际抗癌联盟标准获得了客观反应。当使用CAS评分为0、1 - 100和大于100（反应率分别为0、41%和64%）进行划分时，获得了反应与CAS雌激素受体阳性程度之间的关系。这些数据表明，自动图像分析为免疫细胞化学染色切片中雌激素受体的定量提供了一种可靠、可重复的方法。它相对于手工操作具有潜在优势，在切片评分中受主观影响的机会更少。软件设计的未来进展应进一步减少主观性因素，并提高结果的速度和可靠性。我们预计图像分析将成为例如关于类固醇受体分布异质性对乳房切除术后乳腺癌复发率以及疾病临床进程影响的研究中的一种有价值的工具。
