Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee 247667, India.
Department of Biosciences and Bioengineering, Indian Institute of Technology Roorkee, Roorkee 247667, India.
J Proteomics. 2022 Sep 15;267:104701. doi: 10.1016/j.jprot.2022.104701. Epub 2022 Aug 19.
Triple-negative breast cancer (TNBC) is the most aggressive subtype due to the absence of hormonal receptors. Our study aimed to identify and determine the effectiveness of salivary proteins as candidate markers for metastatic TNBC subtype using parallel reaction monitoring mass spectrometry (PRM-MS). Three salivary proteins (lipocalin-1, SMR3B, and plastin-2) that showed significant differential expression in label-free quantitation (LFQ) between TNBC (N = 6) and health subjects (HS; N = 6) were selected for further validation. The developed PRM assay was used to quantify peptides GLST and NNLE (lipocalin-1), VYAL and MINL (Plastin-2) and GPYP, and IPPP (SMR3B) on a different cohort of TNBC patients (N = 20) and HS (N = 20) for evaluating their discriminating performances. Quantitative validation using PRM correlated well with the LFQ results, and 5 peptides from three proteins showed a similar up-or down-regulation. Subsequently, these proteins were validated by Western blot analysis. Compared to one protein's performance as an individual marker, the five-signature panel with salivary GLST, VYAL, MINL, GPYP, and IPPP achieved better performance in differentiating aggressive TNBC and HS with sensitivity (80%) and specificity (95%). Targeted proteomic analysis of the prioritized proteins highlights a peptide-based signature in saliva as the potential predictor to distinguish between TNBC and HS. SIGNIFICANCE OF THE STUDY: This study was designed to identify and quantify potential markers in saliva from the triple-negative breast cancer (TNBC) patients using parallel reaction monitoring assay. Three salivary proteins, Lipocalin-1 (LCN-1), Submaxillary androgen-regulated protein 3B (SMR3B), and Plastin-2 (LCP-1) selected in the discovery-phase were further quantified by targeted proteomics and Western blots. The salivary proteins successfully differentiated TNBC patients from healthy subjects with a sensitivity (80%) and specificity (95%).
三阴性乳腺癌（TNBC）由于缺乏激素受体，是最具侵袭性的亚型。我们的研究旨在使用平行反应监测质谱法（PRM-MS）鉴定和确定唾液蛋白作为转移性 TNBC 亚型的候选标志物的有效性。在无标记定量（LFQ）中，TNBC（N=6）和健康受试者（HS；N=6）之间显示出显著差异表达的三种唾液蛋白（载脂蛋白 1、SMR3B 和 plastin-2）被选择用于进一步验证。开发的 PRM 测定法用于在不同的 TNBC 患者队列（N=20）和 HS（N=20）上定量肽 GLST 和 NNLE（载脂蛋白 1）、VYAL 和 MINL（Plastin-2）和 GPYP、和 IPPP（SMR3B），以评估它们的区分性能。使用 PRM 进行的定量验证与 LFQ 结果很好地相关，并且来自三种蛋白质的 5 个肽显示出相似的上调或下调。随后，通过 Western blot 分析验证了这些蛋白质。与单个蛋白质作为个体标志物的性能相比，具有唾液 GLST、VYAL、MINL、GPYP 和 IPPP 的五标志物组合在区分侵袭性 TNBC 和 HS 方面表现出更好的性能，具有敏感性（80%）和特异性（95%）。优先蛋白质的靶向蛋白质组学分析突出了唾液中基于肽的特征作为区分 TNBC 和 HS 的潜在预测因子。研究的意义：本研究旨在使用平行反应监测分析从三阴性乳腺癌（TNBC）患者的唾液中鉴定和定量潜在标志物。在发现阶段选择的三种唾液蛋白，载脂蛋白 1（LCN-1）、颌下雄激素调节蛋白 3B（SMR3B）和 Plastin-2（LCP-1），通过靶向蛋白质组学和 Western blot 进一步定量。唾液蛋白成功地将 TNBC 患者与健康受试者区分开来，具有敏感性（80%）和特异性（95%）。
