Influence of decalcification procedures on immunohistochemistry and molecular pathology in breast cancer
Influence of decalcification procedures on immunohistochemistry and molecular pathology in breast cancer
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DOI：
10.1038/modpathol.2016.116
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发表时间：
2016-12-01
期刊：
影响因子：
7.5
通讯作者：
van Diest, Paul J.
中科院分区：
文献类型：
--
作者：
Schrijver, Willemijne A. M. E.;van der Groep, Petra;van Diest, Paul J.
目前，对远处的乳腺癌转移进行常规活组织检查，以重新评估受体状态，并分离DNA用于可用药靶点的测序。骨转移是最常见的亚型。脱钙程序可能会对抗原性和DNA质量产生负面影响。因此，我们评估了几种脱钙程序对受体状态和DNA/RNA质量的影响。在23个前瞻性收集的乳腺肿瘤中，我们通过免疫组织化学方法比较了常规用于诊断目的的(未脱钙的)组织以及在Christensen‘s缓冲液中使用和不使用微波、EDTA和Formical-4的平行脱钙组织中ERα、PR和HER2的状态。此外，还对HER2荧光原位杂交和DNA/RNA的数量和质量进行了评估。结果发现，EDTA(P=0.049)和Formical-4(P=0.047)处理组ERα阳性细胞百分率均低于对照组(P=0.047)，经克里斯滕森缓冲处理后PR表达降低(P=0.041)。总体而言，ERα、PR和HER2免疫组织化学在未脱钙对照组织和脱钙组织中的表达具有良好的一致性(加权kappa)。对于两名患者(9%)，激素治疗或HER2靶向治疗对治疗决策有潜在影响。HER2荧光原位杂交的解释受到Christensen‘s Buffer和Formical-4的严重阻碍，四种脱钙程序都使DNA/RNA的数量和质量下降。在配对的原发乳腺肿瘤标本和EDTA治疗的骨转移瘤标本上的验证表明，免疫组织化学和荧光原位杂交可以很好地评估，DNA和RNA的产量和质量是足够的。有了这一点，我们得出结论，常见的脱钙程序对乳腺癌激素和HER2受体免疫组织化学只有轻微的负面影响。然而，它们可能会严重影响基于DNA/RNA的诊断程序。总体而言，基于EDTA的脱钙是首选的，因为它最能实现荧光原位杂交和DNA/RNA分离。
Distant breast cancer metastases are nowadays routinely biopsied to reassess receptor status and to isolate DNA for sequencing of druggable targets. Bone metastases are the most frequent subgroup. Decalcification procedures may negatively affect antigenicity and DNA quality. We therefore evaluated the effect of several decalcification procedures on receptor status and DNA/RNA quality. In 23 prospectively collected breast tumors, we compared ER alpha, PR and HER2 status by immunohistochemistry in (non-decalcified) tissue routinely processed for diagnostic purposes and in parallel tissue decalcified in Christensen's buffer with and without microwave, EDTA and Formical-4. Furthermore, HER2 fluorescence in situ hybridization and DNA/RNA quantity and quality were assessed. We found that the percentage of ER alpha-positive cells were on average lower in EDTA (P= 0.049) and Formical-4 (P= 0.047) treated cases, compared with controls, and PR expression showed decreased antigenicity after Christensen's buffer treatment (P=0.041). Overall, a good concordance (weighted kappa) was seen for ER alpha, PR and HER2 immunohistochemistry when comparing the non-decalcified control tissues with the decalcified tissues. For two patients (9%), there was a potential influence on therapeutic decision making with regard to hormonal therapy or HER2-targeted therapy. HER2 fluorescence in situ hybridization interpretation was seriously hampered by Christensen's buffer and Formical-4, and DNA/RNA quantity and quality were decreased after all four decalcification procedures. Validation on paired primary breast tumor specimens and EDTA-treated bone metastases showed that immunohistochemistry and fluorescence in situ hybridization were well assessable and DNA and RNA yield and quality were sufficient. With this, we conclude that common decalcification procedures have only a modest negative influence on hormone and HER2 receptor immunohistochemistry in breast cancer. However, they may seriously affect DNA/RNA-based diagnostic procedures. Overall, EDTA-based decalcification is therefore to be preferred as it best allows fluorescence in situ hybridization and DNA/RNA isolation.