口腔癌微环境在疾病的发展和进展中起着重要作用，揭示了疾病的异质性。多种细胞和非细胞因子，包括二肽，据报道具有推动肿瘤进展和转移的作用。在肿瘤微环境中，脯氨酰-羟脯氨酰（Pro-Hyp）是一种与纤维化和口腔癌相关的胶原降解产物。然而，检测口腔癌患者指甲中的Pro-Hyp是一种潜在的生物标记物，而我们对Pro-Hyp的生物学相关性的了解非常有限。在这里，作者试图使用一种新颖的自制垂直管凝胶电泳（VTGE）方案来评估口腔癌患者和健康受试者指甲中Pro-Hyp的水平。此外，我们还采用分子对接和分子动力学（MD）模拟来预测Pro-Hyp的生物功能。我们通过SWISS-ADME服务器评估了Pro-Hyp和已知的磷脂酶C-β2（PLC-β2）激活剂m-3M3FBS的ADME特性，例如药物相似性和引物相似性。我们报告称，在多种关键代谢物中，脯氨酰-羟脯氨酰（Pro-Hyp），一种二肽，减少在口腔癌患者的指甲中。分子对接和MD模拟有助于提出脯氨酰-羟脯氨酰（Pro-Hyp）作为磷脂酶C-β2（PLC-β2）的激活剂的潜在作用。脯氨酰-羟脯氨酰（Pro-Hyp）显示出良好的结合亲和力（-7.6 kcal/mol），通过与关键残基（如HIS311、HIS312、VAL641和GLU743）的常规氢键发生特异性相互作用。MD模拟显示与已知的PLC-β2激活剂m-3M3FBS类似，激活剂结合残基和复合物稳定性。脯氨酰-羟脯氨酰（Pro-Hyp）的药物相似性和引物相似性的ADME特性与m-3M3FBS高度相似，甚至更好。这项研究是首次报道口腔癌患者指甲中Pro-Hyp作为代谢物生物标记物的，进一步建议Pro-Hyp的作用是激活PLC-β2作为促肿瘤信号级联的。未来可以探索作为标志物和Pro-Hyp类似物的诊断和治疗方法。版权所有© Bentham Science Publishers；有关任何疑问，请发送电子邮件至epub@benthamscience.net。

The oral cancer microenvironment plays an important role in the development and progression of the disease which depicts the heterogeneous nature of diseases. Several cellular and non-cellular factors, including dipeptides, have been reported to drive tumor progression and metastasis. Among various secreted molecules in the tumor microenvironment, prolylhydroxyproline (Pro-Hyp) is a collagen-degraded product with specific relevance to fibrosis and oral cancer. However, the detection of Pro-Hyp in the nails of oral cancer patients is a potential biomarker, and our understanding of the biological relevance of Pro-Hyp is highly limited.Here, the authors have attempted to use a novel and in-house vertical tube gel electrophoresis (VTGE) protocol to evaluate the level of Pro-Hyp in the nails of oral cancer patients and healthy subjects. Furthermore, we employed molecular docking and molecular dynamics (MD) simulations to predict the biological function of Pro-Hyp. ADME profiles such as the druglikeness and leadlikeness of Pro-Hyp and a known PLC-β2 activator, m-3M3FBS, were evaluated by the SWISS-ADME server.We report that among various key metabolites, Pro-Hyp, a dipeptide, is reduced in the nails of oral cancer patients. Molecular docking and MD simulations helped to suggest the potential role of Pro-Hyp as an activator of Phospholipase C-β2 (PLC-β2). Pro-Hyp displayed good binding affinity (-7.6 kcal/mol) with specific interactions by a conventional hydrogen bond with key residues, such as HIS311, HIS312, VAL641, and GLU743. MD simulations showed that the activator binding residues and stability of complexes are similar to the well-known activator m-3M3FBS of PLC-β2. ADME profiles such as the druglikeness and leadlikeness of Pro-Hyp were found to be highly comparable and even better than those of m-3M3FBS.This study is one of the first reports on Pro-Hyp as a metabolite biomarker in the nails of oral cancer patients. Furthermore, the implications of Pro-Hyp are proposed to activate PLC-β2 as a pro-tumor signaling cascade. In the future, diagnostic and therapeutic approaches may be explored as biomarkers and mimetic of Pro-Hyp.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.