Institute of Biosciences and BioResources, National Research Council (CNR-IBBR), 80131, Napoli, Italy.
Institute of Biostructure and Bioimaging, National Research Council (CNR-IBB), 80134, Napoli, Italy.
Mol Biol Rep. 2021 Feb;48(2):1505-1519. doi: 10.1007/s11033-020-06129-4. Epub 2021 Jan 20.
Serine hydrolases play crucial roles in many physiological and pathophysiological processes and a panel of these enzymes are targets of approved drugs. Despite this, most of the human serine hydrolases remain poorly characterized with respect to their biological functions and substrates and only a limited number of in vivo active inhibitors have been so far identified. Acylpeptide hydrolase (APEH) is a member of the prolyl-oligopeptidase class, with a unique substrate specificity, that has been suggested to have a potential oncogenic role. In this study, a set of peptides was rationally designed from the lead compound SsCEI 4 and in vitro screened for APEH inhibition. Out of these molecules, a dodecapeptide named Ala 3 showed the best inhibitory effects and it was chosen as a candidate for investigating the anti-cancer effects induced by inhibition of APEH in SAOS-2 cell lines. The results clearly demonstrated that Ala 3 markedly reduced cell viability via deregulation of the APEH-proteasome system. Furthermore, flow cytometric analysis revealed that Ala 3 anti-proliferative effects were closely related to the activation of a caspase-dependent apoptotic pathway. Our findings provide further evidence that APEH can play a crucial role in the pathogenesis of cancer, shedding new light on the great potential of this enzyme as an attractive target for the diagnosis and the quest for selective cancer therapies.
丝氨酸水解酶在许多生理和病理生理过程中发挥着关键作用,其中一组酶是已批准药物的靶点。尽管如此,大多数人类丝氨酸水解酶的生物学功能和底物仍未得到充分描述,迄今为止只鉴定出有限数量的体内活性抑制剂。酰肽水解酶 (APEH) 是脯氨酰寡肽酶类的成员,具有独特的底物特异性,据推测其具有潜在的致癌作用。在这项研究中,从先导化合物 SsCEI 4 中合理设计了一组肽,并在体外筛选 APEH 抑制作用。在这些分子中,一种名为 Ala 3 的十二肽显示出最好的抑制效果,因此被选为研究抑制 APEH 在 SAOS-2 细胞系中诱导的抗癌作用的候选物。结果清楚地表明,Ala 3 通过调节 APEH-蛋白酶体系统显著降低细胞活力。此外,流式细胞术分析表明,Ala 3 的抗增殖作用与 caspase 依赖性凋亡途径的激活密切相关。我们的研究结果进一步证明 APEH 在癌症发病机制中起着至关重要的作用,为该酶作为有吸引力的诊断和选择性癌症治疗目标提供了新的证据。