实验动物与比较医学 ›› 2020, Vol. 40 ›› Issue (5): 426-.DOI: 10.3969/j.issn.1674-5817.2020.05.011

• 论著 • 上一篇    下一篇

基于冷冻肿瘤组织切片的磷酸化HER2免疫组织化学染色方法探讨

范姝琼1, 2,郑莉1   

  1. 1. 迪哲(上海)医药有限公司,上海 201203; 2. 复旦大学生命科学学院,上海 200433
  • 收稿日期:2019-09-12 出版日期:2020-10-15 发布日期:2020-11-19
  • 作者简介:范姝琼(1988—), 女, 硕士研究生, 主要从事肿瘤分子病理研究。E-mail: shin0323@163.com

Exploration of Phospho-HER2 Immunohistochemistry Staining on Frozen Tumor Tissues

FAN Shuqiong1,2, ZHENG Li1   

  1. 1. Dizal (Shanghai) Pharmaceutical Co.,Ltd, Shanghai 201203, China; 
    2. School of Life Sciences, Fudan University, Shanghai 200433, China
  • Received:2019-09-12 Online:2020-10-15 Published:2020-11-19

摘要: 目的   探索针对磷酸化人类表皮生长因子受体2(human epidermal growth factor receptor 2,HER2)免疫组织化学染色实验中冷冻组织切片的最佳固定与保存条件。方法    在BALB/c-nu/nu小鼠中建立人源性乳腺癌细胞BT474的皮下移植瘤模型,对比不同的抗原固定条件和存储时间下,免疫组织化学染色观察肿瘤模型中磷酸化HER2的信号变化。结果   冰冻切片用体积分数为10%的甲醛溶液固定5 min,可以维持良好的细胞形态。在-20 ℃的存储条件下,肿瘤组织中磷酸化HER2蛋白在12周内表达稳定。结论    冰冻组织切片进行免疫组织化学染色用时短,可以作为常规石蜡切片染色方法的补充或者替代。

关键词: 冷冻切片, 人类表皮生长因子受体2, 免疫组织化学, BALB/c-nu/nu小鼠

Abstract: Objective    To investigate the optimized fixation and storage conditions for the immunohistochemistry detection of phospho-human epidermal growth factor receptor 2 (HER2) on frozen sections. Methods    Xenograft model was established by subcutaneously injecting human breast cancer cell BT474 into the abdomen of BALB/c-nu/nu mice. The mice were sacrificed when the tumor volume up to 300 mm3. The tumor samples were quickly frozen, sectioned, fixed in different reagents and stored for different time period. Phospho-HER2 immunohistochemistry was performed to evaluate the change of protein expression under these conditions. Results    Good cell morphology was kept by the fixation of frozen section in 50% formalin for 5 minutes. The expression of phospho-HER2 protein was stable within 12 weeks after fixation when the tumor tissues were stored in -20 ℃. Conclusion   The immunohistochemistry staining on frozen tissues takes less time and can be a supplement or replacement of traditional staining on formalin-fixed and paraffin-embedded sections. 


Key words: Frozen section, Human epidermal growth factor receptor 2, Immunohistochemistry; BALB/c-nu/nu mice

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