Wei Yali, An Xu, Cao Qinmei, Che Nanying, Xue Yuanyuan, Deng Haiteng, Wang Qingtao, Zhou Rui
Department of Clinical Laboratory, Beijing Chaoyang Hospital, The Third Clinical Medical College of Capital Medical University, Beijing, China.
Department of Clinical Laboratory, Tongzhou Maternal and Child Health Hospital of Beijing, Beijing, China.
Front Oncol. 2022 Aug 16;12:955943. doi: 10.3389/fonc.2022.955943. eCollection 2022.
The human epidermal growth factor receptor 2 (HER2) is an important biomarker that plays a pivotal role in therapeutic decision-making for patients with breast cancer (BC). Patients with HER2-low BC can benefit from new HER2 targeted therapy. For ensuring the accurate and reproducible detection of HER2-low cancer, reliable reference materials are required for monitoring the sensitivity and specificity of detection assays. Herein, a lentiviral vector was used to transduce the HER2 gene into MDA-MB-231 cells that exhibited low HER2 density, and the cells were characterized by droplet digital PCR to accurately determine the copy number variation. Then, the formalin-fixed paraffin-embedded (FFPE) samples from xenografts were prepared and evaluated for suitability as candidate reference materials by immunohistochemistry (IHC) and fluorescence hybridization (FISH). The FFPE reference materials were selected on the basis of IHC score of 2+ and negative FISH result to meet the requirement for HER2-low BC detection. Furthermore, the FFPE reference materials exhibited typical histological structures that resembled the clinical BC specimens. These novel FFPE reference materials displayed the high stability and homogeneity, and they were produced in high quantity. In summary, we generated high-quality reference materials for internal quality control and proficiency testing in HER2-low detection.
人表皮生长因子受体2（HER2）是一种重要的生物标志物，在乳腺癌（BC）患者的治疗决策中起着关键作用。HER2低表达的BC患者可从新的HER2靶向治疗中获益。为确保准确、可重复地检测HER2低表达癌症，需要可靠的参考材料来监测检测方法的灵敏度和特异性。在此，使用慢病毒载体将HER2基因转导至HER2密度低的MDA-MB-231细胞中，并通过液滴数字PCR对细胞进行表征，以准确确定拷贝数变异。然后，制备来自异种移植瘤的福尔马林固定石蜡包埋（FFPE）样本，并通过免疫组织化学（IHC）和荧光杂交（FISH）评估其作为候选参考材料的适用性。根据IHC评分为2+且FISH结果为阴性来选择FFPE参考材料，以满足HER2低表达BC检测的要求。此外，FFPE参考材料呈现出与临床BC标本相似的典型组织结构。这些新型FFPE参考材料具有高稳定性和均一性，且产量高。总之，我们生成了用于HER2低表达检测内部质量控制和能力验证的高质量参考材料。
