Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, Via Vetoio, Coppito 2, 67100, L'Aquila, Italy.
Expert Rev Proteomics. 2013 Jun;10(3):223-6. doi: 10.1586/epr.13.18.
Wulfkuhle JD, Berg D, Wolff C et al. Molecular analysis of HER2 signaling in human breast cancer by functional protein pathway activation mapping. Clin. Cancer Res. 18(23), 6426-6435 (2012). Exhaustive characterization and mapping of pivotal molecules and downstream effectors deregulated in breast cancer is of fundamental clinical value to define the most effective therapy. Wulfkuhle et al. applied reverse-phase protein microarray, a highly sensitive immunoassay able to perform quantitative and multiplexed analysis of total and/or modified cellular proteins, to assess protein levels and activation/phosphorylation status of the HER family (EGFR, HER2, HER3) and downstream signaling molecules in HER2(+) and HER2(-) breast cancers. The research was performed using laser capture microdissected tumor epithelial cells from frozen samples and formalin-fixed paraffin embedded specimens, which were also analyzed by immunohistochemistry (IHC) and FISH. This study identified a subgroup of IHC/FISH/HER2(-) patients with HER2 activation/phosphorylation levels comparable with those obtained from IHC/FISH/HER2(+) tumors. HER2 signaling activation was independent from total HER2 expression and involved HER3 and EGFR activation. These findings indicate that molecular characterization by reverse-phase protein microarray of HER2 and its partners/effectors in the signaling cascade enables the identification of a subgroup of IHC/FISH/HER2(-) patients showing HER2 signaling activation. These patients, currently excluded from targeted therapy administration, could potentially benefit from this and it could improve prognosis and survival.
Wulfkuhle JD、Berg D、Wolff C 等人。通过功能蛋白通路激活图谱对人类乳腺癌中 HER2 信号的分子分析。临床癌症研究。18(23)，6426-6435(2012)。彻底描述和绘制乳腺癌中关键分子和下游效应物的失调图谱，对于定义最有效的治疗方法具有重要的临床价值。Wulfkuhle 等人应用反相蛋白质微阵列，这是一种高度敏感的免疫测定法，能够对总细胞和/或修饰细胞蛋白进行定量和多重分析，以评估 HER 家族（EGFR、HER2、HER3）和下游信号分子在 HER2(+)和 HER2(-)乳腺癌中的蛋白水平和激活/磷酸化状态。该研究使用来自冷冻样本和福尔马林固定石蜡包埋标本的激光捕获微切割肿瘤上皮细胞进行，这些标本也通过免疫组织化学（IHC）和 FISH 进行分析。这项研究确定了一个 IHC/FISH/HER2(-)亚组患者，其 HER2 激活/磷酸化水平与从 IHC/FISH/HER2(+)肿瘤中获得的水平相当。HER2 信号激活独立于总 HER2 表达，涉及 HER3 和 EGFR 激活。这些发现表明，通过反相蛋白质微阵列对 HER2 及其在信号级联中的伙伴/效应物进行分子特征分析，可以识别出一组 IHC/FISH/HER2(-)患者，这些患者表现出 HER2 信号激活。这些目前被排除在靶向治疗管理之外的患者可能从中受益，这可以改善预后和生存。
