DataSheet1_Integrated analysis of Dendrobium nobile extract Dendrobin A against pancreatic ductal adenocarcinoma based on network pharmacology, bioinformatics, and validation experiments.PDF

Background:Dendrobium nobile (D. nobile), a traditional Chinese medicine, has received attention as an anti-tumor drug, but its mechanism is still unclear. In this study, we applied network pharmacology, bioinformatics, and in vitro experiments to explore the effect and mechanism of Dendrobin A, the active ingredient of D. nobile, against pancreatic ductal adenocarcinoma (PDAC).
Methods: The databases of SwissTargetPrediction and PharmMapper were used to obtain the potential targets of Dendrobin A, and the differentially expressed genes (DEGs) between PDAC and normal pancreatic tissues were obtained from The Cancer Genome Atlas and Genotype-Tissue Expression databases. The protein-protein interaction (PPI) network for Dendrobin A anti-PDAC targets was constructed based on the STRING database. Molecular docking was used to assess Dendrobin A anti-PDAC targets. PLAU, one of the key targets of Dendrobin A anti-PDAC, was immunohistochemically stained in clinical tissue arrays. Finally, in vitro experiments were used to validate the effects of Dendrobin A on PLAU expression and the proliferation, apoptosis, cell cycle, migration, and invasion of PDAC cells.
Results: A total of 90 genes for Dendrobin A anti-PDAC were screened, and a PPI network for Dendrobin A anti-PDAC targets was constructed. Notably, a scale-free module with 19 genes in the PPI indicated that the PPI is highly credible. Among these 19 genes, PLAU was positively correlated with the cachexia status while negatively correlated with the overall survival of PDAC patients. Through molecular docking, Dendrobin A was found to bind to PLAU, and the Dendrobin A treatment led to an attenuated PLAU expression in PDAC cells. Based on clinical tissue arrays, PLAU protein was highly expressed in PDAC cells compared to normal controls, and PLAU protein levels were associated with the differentiation and lymph node metastatic status of PDAC. In vitro experiments further showed that Dendrobin A treatment significantly inhibited the proliferation, migration, and invasion, inducing apoptosis and arresting the cell cycle of PDAC cells at the G2/M phase.
Conclusion: Dendrobin A, a representative active ingredient of D. nobile, can effectively fight against PDAC by targeting PLAU. Our results provide the foundation for future PDAC treatment based on D. nobile.

背景：石斛（Dendrobium nobile, D. nobile）作为传统中药材，其作为抗肿瘤药物已受到广泛关注，但具体作用机制仍未阐明。本研究运用网络药理学、生物信息学及体外实验，探索石斛活性成分石斛碱A（Dendrobin A）对抗胰腺导管腺癌（pancreatic ductal adenocarcinoma, PDAC）的作用及潜在机制。
方法：本研究通过SwissTargetPrediction与PharmMapper数据库获取石斛碱A的潜在靶点；从癌症基因组图谱（The Cancer Genome Atlas）及基因型组织表达数据库（Genotype-Tissue Expression）中获取胰腺导管腺癌与正常胰腺组织的差异表达基因（differentially expressed genes, DEGs）。基于STRING数据库构建石斛碱A抗胰腺导管腺癌靶点的蛋白质相互作用（protein-protein interaction, PPI）网络；采用分子对接技术评估石斛碱A与抗胰腺导管腺癌靶点的结合活性。对临床组织芯片中的关键靶点PLAU（尿激酶型纤溶酶原激活物）进行免疫组化染色。最后通过体外实验验证石斛碱A对PDAC细胞中PLAU表达水平，以及细胞增殖、凋亡、细胞周期、迁移与侵袭的影响。
结果：本研究共筛选得到90个与石斛碱A抗胰腺导管腺癌作用相关的基因，并构建了对应的PPI网络。值得注意的是，该PPI网络中存在一个包含19个基因的无尺度模块，提示该网络具有较高可信度。在这19个基因中，PLAU与胰腺导管腺癌患者的恶病质状态呈正相关，而与患者总生存期呈负相关。分子对接结果显示，石斛碱A可与PLAU特异性结合；且石斛碱A处理可显著下调PDAC细胞中PLAU的表达水平。基于临床组织芯片的分析表明，相较于正常对照组织，PLAU蛋白在PDAC细胞中呈高表达，且PLAU蛋白水平与PDAC的分化程度及淋巴结转移状态密切相关。体外实验进一步证实，石斛碱A处理可显著抑制PDAC细胞的增殖、迁移与侵袭，诱导细胞凋亡，并将PDAC细胞周期阻滞于G2/M期。
结论：作为石斛的代表性活性成分，石斛碱A可通过靶向PLAU有效对抗胰腺导管腺癌。本研究结果为基于石斛的胰腺导管腺癌治疗研究奠定了理论基础。