摘要目的 探讨硝酸、乙二胺四乙酸(EDTA)和RapidCal三种不同脱钙液在乳腺癌骨转移标本的免疫组织化学和HER2荧光原位杂交(FISH)检测中的表现.方法 选取28例乳腺癌骨转移标本,建立统一的前处理方案,分别用不同的脱钙液(10%硝酸、10%EDTA、RapidCal脱钙液)进行分组脱钙,观察HE染色和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)、HER2等免疫组织化学检测及HER2 FISH结果,并进行比较及评估.结果 HE染色结果表明三组形态学无明显差异.RapidCal组对免疫组织化学各类定位的抗原保存均完好,具有和标准脱钙液EDTA相似的染色效果明显优于硝酸组(P<0.05).硝酸组核计数缺失严重,Ki-67几乎为0,胞质抗原(RANK、RANKL)及胞膜抗原(HER2)丢失幅度小于核抗原.RapidCal组与EDTA组FISH结果一致,且与HER2免疫组织化学结果相符合,而硝酸组HER2蛋白表达信号明显下降,第17号染色体信号缺失(P<0.05).结论 硝酸脱钙液处理后的标本染色效果较差,新型进口脱钙剂RapidCal和EDTA脱钙液处理的乳腺癌骨转移标本,在形态学、常用免疫组织化学和FISH的应用上表现出优秀的标本质量,但由于EDTA脱钙液脱钙时间过长,使用新型进口脱钙剂RapidCal将有利于质量控制及临床推广.

abstractsObjective To investigate the optimal strategy for immunohistochemical (IHC) staining in bone metastasis specimens from breast cancer.Methods 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.Results 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 (P<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 (P<0.05).Conclusions 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.