Abstract：
Transmembrane protease/serine (TMPRSS2), a type II transmembrane serine protease, plays a crucial role in different stages of cancer. Recent studies have reported that the triggering epidermal growth factor receptor (EGFR) activation through protease action promotes metastasis. However, there are no reports on the interaction of TMPRSS2 with EGFR, especially in triple-negative triple negative (TNBC). The current study investigates the unexplored interaction between TMPRSS2 and EGFR, which are key partners mediating metastasis. This interaction is explored for potential targeting using quercetin (QUE) and taxifolin (TAX). TMPRSS2 expression patterns in breast cancer (BC) tissues and subtypes have been predicted, with the prognostic significance assessed using the GENT2.0 database. Validation of TMPRSS2 expression was performed in normal and TNBC tissues, including drug-resistant cell lines, utilizing GEO datasets. TMPRSS2 was further validated as a predictive biomarker for FDA-approved chemotherapeutics through transcriptomic data from BC patients. The study demonstrated the association of TMPRSS2 with EGFR through in silico analysis and validates the findings in TNBC cohorts using the TIMER2.0 web server and the TCGA dataset through C-Bioportal. Molecular docking and molecular dynamic simulation studies identified QUE and TAX as best leads targeting TMPRSS2. They inhibited cell-free TMPRSS2 activity like clinical inhibitor of TMPRSS2, Camostat mesylate. In cell-based assays focused on paclitaxel-resistant TNBC (TNBC/PR), QUE and TAX demonstrated potent inhibitory activity against extracellular and membrane-bound TMPRSS2, with low IC50 values. Furthermore, ELISA and cell-based AlphaLISA assays demonstrated that QUE and TAX inhibit the interaction of TMPRSS2 with EGFR. Additionally, QUE and TAX exhibited significant inhibition of proliferation and cell cycle accompanied by notable alterations in the morphology of TNBC/PR cells. This study provides valuable insights into potential of QUE and TAX targeting TMPRSS2 overexpressing TNBC.
摘要：
跨膜蛋白酶/丝氨酸（TMPRSS2），一种II型跨膜丝氨酸蛋白酶,在癌症的不同阶段起着至关重要的作用。最近的研究报道，通过蛋白酶作用触发表皮生长因子受体（EGFR）激活促进转移。然而,没有关于TMPRSS2与EGFR相互作用的报道，特别是在三负三负（TNBC）。目前的研究调查了TMPRSS2和EGFR之间尚未探索的相互作用，它们是介导转移的关键伙伴。使用槲皮素（QUE）和紫杉素（TAX）探索这种相互作用的潜在靶向性。TMPRSS2在乳腺癌（BC）组织和亚型中的表达模式已被预测，使用GENT2.0数据库评估预后意义。在正常组织和TNBC组织中进行TMPRSS2表达的验证，包括耐药细胞系，利用GEO数据集。通过来自BC患者的转录组数据,TMPRSS2被进一步验证为FDA批准的化疗药物的预测性生物标志物。该研究通过计算机分析证明了TMPRSS2与EGFR的关联，并通过C-Bioportal使用TIMER2.0Web服务器和TCGA数据集验证了TNBC队列中的发现。分子对接和分子动态模拟研究确定QUE和TAX是靶向TMPRSS2的最佳线索。它们抑制无细胞TMPRSS2活性，就像TMPRSS2的临床抑制剂，甲磺酸卡莫司他一样。在集中于紫杉醇抗性TNBC（TNBC/PR）的基于细胞的测定中，QUE和TAX表现出对细胞外和膜结合的TMPRSS2的有效抑制活性,具有低IC50值。此外，ELISA和基于细胞的AlphaLISA测定证明QUE和TAX抑制TMPRSS2与EGFR的相互作用。此外,QUE和TAX表现出对增殖和细胞周期的显著抑制,伴随着TNBC/PR细胞形态的显著改变。这项研究为QUE和TAX靶向TMPRSS2过表达TNBC的潜力提供了有价值的见解。
