Examining the technique of angiogenesis assessment in invasive breast cancer.
Examining the technique of angiogenesis assessment in invasive breast cancer.
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DOI：
10.1038/bjc.1997.506
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发表时间：
1997
影响因子：
8.8
通讯作者：
Winstanley J
中科院分区：
文献类型：
--
作者：
Martin L;Green B;Renshaw C;Lowe D;Rudland P;Leinster SJ;Winstanley J
据报道，在一些（但不是全部）研究中，通过微血管密度测量的血管生成强度与浸润性乳腺癌的不良预后相关。造成这些差异的原因可能是所使用的方法的差异。用于识别微血管的单克隆抗体、计数的高密度区域或“热点”的数量以及用于统计分析的值的类型（最高计数或平均计数）在不同的研究之间有所不同。我们评估了三种常用单克隆抗体中的哪一种能够提供浸润性乳腺癌中微血管的最佳可视化效果，并使用一些方法为每种试剂的最佳“热点”数量提供可重复的数据。因此，使用针对 FVIIIRAg、CD31 和 CD34 的单克隆抗体对来自 174 个原发性乳腺癌的福尔马林固定石蜡包埋标本中的微血管进行免疫组织化学染色，并对每种抗体在 200 倍放大倍数下计数 10 个视野。采用最高计数以及3次、5次和10次计数中最高计数的平均值来考察微血管密度与中位随访时间为7.1年的患者总生存率之间的关系。 CD31 和 CD34 抗体比 FVIIIRAg 抗体识别出更多血管（每 mm2 中位最高计数：CD31 = 100、CD34 = 100、FVIIIRAg = 81）。然而，针对 CD31 的单克隆抗体是最不可靠的抗体，仅对 87% 的切片进行免疫组织化学染色，而针对 CD34 的单克隆抗体为 98%，针对 FVIIIRAg 的单克隆抗体为 99%。不同抗体染色的血管数量之间存在高度相关性，尽管同一标本的连续切片的不同抗体染色的实际计数存在相当大的差异。患者可分为两组，分别对应于微血管密度高的组和微血管密度低的组。使用 Kaplan-Meier 生存曲线，无论使用哪种记录最高血管密度的方法，所有三种抗体在血管密度和生存之间都存在密切关联。使用对数秩检验和 Cox 回归分析，抗 CD34 给出了三种抗体中最显着的结果，而高组和低组的第 75 个百分位数的简单截止值与患者生存率产生了最佳关联。对于抗CD34，最高微血管密度（P = 0.0014）和最高三个微血管密度的平均值（P = 0.004）显示与患者死亡具有良好的相关性，而对于抗CD31（P = 0.008）和抗FVIIIRAg（P = 0.007），使用Cox回归分析，最高计数给出了最佳相关性。
The intensity of angiogenesis as measured by the density of microvessels has been reported to be associated with a poor prognosis in invasive breast cancer in some, but not all, studies. The reasons for these discrepancies may be variations in the methodologies used. The monoclonal antibody used to identify the microvessels, the number of high-density areas or 'hotspots' counted and the type of value taken for statistical analysis (highest count or mean count) have varied between the different studies. We have assessed which of the three commonly used monoclonal antibodies provides the best visualization of microvessels in invasive breast cancer and have used methods that give reproducible data for the optimum number of 'hotspots' to count for each reagent. Thus, microvessels in formalin-fixed paraffin-embedded specimens from 174 primary breast cancers were immunohistochemically stained with monoclonal antibodies to FVIIIRAg, CD31 and CD34 and ten fields counted at 200 x magnification for each antibody. The highest count and the mean value of the highest of three, five and ten counts were used to examine the relationship between the density of microvessels and overall survival of patients with a median follow-up time of 7.1 years. Antibodies to CD31 and CD34 identified more vessels than antibodies to FVIIIRAg (median highest count per mm2: CD31 = 100, CD34 = 100, FVIIIRAg = 81). The monoclonal antibody to CD31, however, was the least reliable antibody, immunohistochemically staining only 87% of sections compared with 98% for the monoclonal to CD34 and 99% for the monoclonal to FVIIIRAg. There was a high degree of correlation between the number of vessels stained by the different antibodies, though there were some considerable differences in actual counts for serial sections of the same specimen stained by the different antibodies. Patients could be divided into two groups corresponding to those with high microvessel densities and those with low microvessel densities. Using Kaplan-Meier survival curves, there was a close association for all three antibodies between vessel density and survival whichever method of recording the highest vessel densities was used. Using log-rank tests and Cox's regression analysis, anti-CD34 gave the most significant results of the three antibodies, whereas a simple cut-off at the 75th percentile for the high and low groups produced the best association with patient survival. For anti-CD34 the highest microvessel density (P = 0.0014) and the mean value of the highest three microvessel densities (P = 0.004) showed a good correlation with patient death, whereas for anti-CD31 (P = 0.008) and anti-FVIIIRAg (P = 0.007) the highest count gave the best correlation using Cox's regression analysis.