Data_Sheet_1_An Advanced Systems Pharmacology Strategy Reveals AKR1B1, MMP2, PTGER3 as Key Genes in the Competing Endogenous RNA Network of Compound Kushen Injection Treating Gastric Carcinoma by Integrated Bioinformatics and Experimental Verification.docx

Gastric carcinoma (GC) is a severe tumor of the digestive tract with high morbidity and mortality and poor prognosis, for which novel treatment options are urgently needed. Compound Kushen injection (CKI), a classical injection of Chinese medicine, has been widely used to treat various tumors in clinical practice for decades. In recent years, a growing number of studies have confirmed that CKI has a beneficial therapeutic effect on GC, However, there are few reports on the potential molecular mechanism of action. Here, using systems pharmacology combined with proteomics analysis as a core concept, we identified the ceRNA network, key targets and signaling pathways regulated by CKI in the treatment of GC. To further explore the role of these key targets in the development of GC, we performed a meta-analysis to compare the expression differences between GC and normal gastric mucosa tissues. Functional enrichment analysis was further used to understand the biological pathways significantly regulated by the key genes. In addition, we determined the significance of the key genes in the prognosis of GC by survival analysis and immune infiltration analysis. Finally, molecular docking simulation was performed to verify the combination of CKI components and key targets. The anti-gastric cancer effect of CKI and its key targets was verified by in vivo and in vitro experiments. The analysis of ceRNA network of CKI on GC revealed that the potential molecular mechanism of CKI can regulate PI3K/AKT and Toll-like receptor signaling pathways by interfering with hub genes such as AKR1B1, MMP2 and PTGERR3. In conclusion, this study not only partially highlighted the molecular mechanism of CKI in GC therapy but also provided a novel and advanced systems pharmacology strategy to explore the mechanisms of traditional Chinese medicine formulations.

胃癌（Gastric carcinoma, GC）是一种高发、高致死且预后不佳的消化道恶性肿瘤，临床上亟需探索全新的治疗方案。复方苦参注射液（Compound Kushen Injection, CKI）作为经典中药注射剂，在临床中已被广泛用于多种肿瘤的治疗长达数十年之久。近年来，越来越多的研究证实复方苦参注射液对胃癌具有良好的治疗效果，但针对其潜在分子作用机制的相关报道却较为匮乏。本研究以系统药理学结合蛋白质组学分析为核心思路，筛选并鉴定出复方苦参注射液治疗胃癌过程中所调控的ceRNA（competing endogenous RNA）网络、关键靶点及信号通路。为进一步明确上述关键靶点在胃癌发生发展中的作用，本研究通过荟萃分析对比了胃癌组织与正常胃黏膜组织的基因表达差异。随后通过功能富集分析，解析关键基因所显著调控的生物学通路。此外，本研究还通过生存分析与免疫浸润分析，明确了关键基因在胃癌预后评估中的临床价值。最后通过分子对接模拟实验验证了复方苦参注射液活性成分与关键靶点的结合活性，并通过体内外实验验证了复方苦参注射液及其关键靶点的抗胃癌活性。针对复方苦参注射液治疗胃癌的ceRNA网络分析结果显示，其潜在分子机制可通过调控AKR1B1、MMP2及PTGERR3等核心靶点，干预PI3K/AKT及Toll样受体信号通路。综上，本研究不仅部分阐明了复方苦参注射液治疗胃癌的分子机制，同时也为探索中药复方的作用机制提供了一种新颖且先进的系统药理学研究策略。