穿山甲是一种名贵中药，在治疗乳腺相关疾病方面有着悠久的药用历史。但由于资源的过度开发利用，穿山甲已被列入珍稀濒危野生动物名录。穿山甲资源的节约和持续临床应用已成为迫切需要解决的问题，而寻找穿山甲的小分子替代品是实现这一目标的有效途径。先前的研究已确定 PA3264 可能是穿山甲中的活性成分。在本研究中，我们系统地研究了PA3264在治疗具有代表性的乳腺相关疾病三阴性乳腺癌（TNBC）中的药理作用和机制。体外。接下来，建立4T1原位肿瘤和人源化PBMC-CDX小鼠模型来检查PA3264的体内抗肿瘤作用。进行转录组测序和分子对接实验来预测功能途径。采用Western blotting和实时定量PCR验证PA3264抗癌作用的分子机制。PA3264在体外显着抑制乳腺癌细胞的细胞活力和迁移。此外，PA3264 抑制了体内肿瘤的大小并减轻了肿瘤的重量。最后证实PA3264通过抑制PI3K/AKT/NF-κB通路，引起细胞周期阻滞，促进细胞凋亡来阻止乳腺癌的进展。本研究阐明，源自珍稀濒危穿山甲的PA3264是一种新型生物活性物质。从科学研究的角度治疗三阴性乳腺癌的肽。© 2024。作者。

Squama Manis is a valuable traditional Chinese medicine with a long history of medicinal use in the treatment of breast-related diseases. However, owing to the excessive exploitation and utilization of the resources, Squama Manis has been included in the list of rare and endangered wild animals. The conservation of the resources of Squama Manis and continuing its clinical application has become an urgent problem, and the search for small-molecule substitutes for Squama Manis is an effective way to achieve this goal. Previous studies have identified PA3264 as a possible active ingredient in Squama Manis. In this study, we systematically investigated the pharmacological effects and mechanisms of PA3264 in the treatment of triple-negative breast cancer (TNBC), a representative breast-related disease.Cell viability and colony formation assays were performed after treatment with the target dipeptide PA3264 in vitro. Next, 4T1 orthotopic tumors and humanized PBMC-CDX mouse models were generated to examine the antitumor effect of PA3264 in vivo. Transcriptome sequencing and molecular docking experiments were performed to predict pathways to function. Western blotting and quantitative real-time PCR were used to validate the molecular mechanisms underlying the anticancer effects of PA3264.PA3264 significantly inhibited cell viability and migration of breast cancer cells in vitro. Furthermore, PA3264 suppressed the tumor size and reduced the tumor weight in vivo. Finally, it was verified that PA3264 prevented the progression of breast cancer by inhibiting the PI3K/AKT/NF-κB pathway, causing cell cycle arrest, and promoting apoptosis.This study elucidated that PA3264 derived from rare and endangered Squama Manis was a novel bioactive peptide for treating triple-negative breast cancer from a scientific research perspective.© 2024. The Author(s).