Wu H Y, Wang T, Chen K L, Fan Z W, Pu X H, Zhang B, Fan X S, Meng F Q
Department of Pathology, the Affiliated Drum Tower Hospital, Nanjing University Medical School, Nanjing 210008, China.
Zhonghua Bing Li Xue Za Zhi. 2017 Aug 8;46(8):536-541. doi: 10.3760/cma.j.issn.0529-5807.2017.08.004.
To investigate the optimal strategy for immunohistochemical (IHC) staining in bone metastasis specimens from breast cancer. Twenty-eight bone metastases specimens from breast cancers were divided into three groups and subjected to different decalcifying agents (group A-10% nitrate, group B-EDTA decalcification, and group C-imported decalcifying solution RapidCal). The effects of those on HE and IHC staining for Ki-67, ER, PR, GATA3, RANK, RANKL, HER2 and HER2 FISH results were assessed. There were no significant differences among three groups in HE morphology and IHC staining. Antigen content in the RapidCal group were all intact; the EDTA group showed a similar staining rate, which was better than the nitrate group (<0.05). Nitrate group showed marked reduction in nuclear Ki-67 staining, but the loss of cytoplasmic antigens (RANK, RANKL) was less than cell membrane antigen (HER2). For FISH, the RapidCal group and EDTA group showed same results, concordant with IHC staining results. The expression of HER2 protein in the nitric acid group was significantly decreased and chromosome 17 labelling was lost (<0.05). RapidCal treated bone metastases specimens from breast cancer show excellent sample quality in morphological, IHC and FISH results compared with traditional decalcifying agents. Owing to the longer time of EDTA decalcification, the new decalcifying agent RapidCal plays an important role in quality control and clinical application.
探讨乳腺癌骨转移标本免疫组织化学（IHC）染色的最佳策略。将28例乳腺癌骨转移标本分为三组，分别使用不同的脱钙剂（A组-10%硝酸，B组-乙二胺四乙酸（EDTA）脱钙，C组-进口脱钙液RapidCal）。评估它们对苏木精-伊红（HE）染色以及Ki-67、雌激素受体（ER）、孕激素受体（PR）、GATA3、核因子κB受体活化因子（RANK）、核因子κB受体活化因子配体（RANKL）、人表皮生长因子受体2（HER2）的IHC染色及HER2荧光原位杂交（FISH）结果的影响。三组在HE形态和IHC染色方面无显著差异。RapidCal组的抗原含量均完整；EDTA组的染色率相似，优于硝酸组（<0.05）。硝酸组细胞核Ki-67染色明显减少，但细胞质抗原（RANK、RANKL）的丢失少于细胞膜抗原（HER2）。对于FISH，RapidCal组和EDTA组结果相同，与IHC染色结果一致。硝酸组中HER2蛋白表达显著降低，17号染色体标记丢失（<0.05）。与传统脱钙剂相比，RapidCal处理的乳腺癌骨转移标本在形态学、IHC和FISH结果方面显示出优异的样本质量。由于EDTA脱钙时间较长，新型脱钙剂RapidCal在质量控制和临床应用中发挥着重要作用。
