Table_1_An Integrative Serum Pharmacology-Based Approach to Study the Anti-Tumor Activity of B. paniculatum Aqueous Bulb Extract on the Human Hepatocellular Carcinoma Cell Line BEL-7404.xlsx

The herb Bolbostemma paniculatum (Maxim) Franquet (Cucurbitaceae family), also known as Tu-Bei-Mu (TBM) in Chinese, has shown curative effects to treat several types of cancer as an adjunctive therapy. Thereby we intend to find its effect on the human hepatocellular carcinoma (HCC) and to understand the pharmacological mechanism behind it. In this study, an integrative serum pharmacology-based approach linking serum pharmacology and bioinformatics prediction was employed. Firstly, we used the serum taken introgastrically from the rats dministered by TBM aqueous bulb extract to culture the HCC cell line BEL-7404 and detect its anti-tumor effects. Secondly, the TBM putative targets were predicted using the ETCM database and known therapeutic targets of NPC were collected from the OMIM database. Then, a TBM-HCC putative targets network was constructed using the DAVID and STRING databases. Thirdly, key gene targets were obtained based on topological analysis and pathway enrichment analysis. The expression of 4 representative key targets were validated by Western blotting. As a result, 36 TBM targets and 26 known therapeutic targets of HCC were identified. These key targets were found to be frequently involved in 13 KEGG pathways and 4 biological processes. The expression of four representative key targets: TP53, CASP3, BCL2 and BAX further supports the suppression of TBM on HCC. In general, our study shows the curative effects of TBM against HCC. By using this integrative approach, we may find novel potential therapeutic targets to suppress HCC using TBM as an adjunctive therapy. And it could also help us understand the mechanism of HCC treatments in response to TBM.

葫芦科土贝母属植物Bolbostemma paniculatum (Maxim) Franquet，其中文名为土贝母（Tu-Bei-Mu, TBM），作为辅助疗法对多种癌症已展现出确切治疗功效。本研究旨在探究其对人肝细胞癌（human hepatocellular carcinoma, HCC）的作用，并阐明其背后的药理机制。本研究采用了一种整合血清药理学与生物信息学预测的综合研究策略。首先，我们使用土贝母鳞茎水提物灌胃大鼠后采集的血清，培养人肝癌细胞系BEL-7404并检测其抗肿瘤活性；其次，采用ETCM数据库（Encyclopedia of Traditional Chinese Medicine）预测土贝母的潜在靶点，并从OMIM数据库（Online Mendelian Inheritance in Man）中获取NPC的已知治疗靶点，随后借助DAVID数据库（Database for Annotation, Visualization and Integrated Discovery）与STRING数据库（Search Tool for the Retrieval of Interacting Genes）构建土贝母-肝细胞癌潜在靶点调控网络；第三，通过拓扑分析与通路富集分析筛选得到核心基因靶点，并采用蛋白质印迹（Western blotting）验证4个代表性核心靶点的表达水平。本研究共鉴定出36个土贝母靶点与26个肝细胞癌已知治疗靶点，上述核心靶点显著富集于13条KEGG（Kyoto Encyclopedia of Genes and Genomes）通路及4个生物学过程。TP53、CASP3、BCL2及BAX这4个代表性核心靶点的表达结果进一步证实，土贝母可抑制肝细胞癌的进展。综上，本研究证实土贝母对肝细胞癌具有明确治疗作用。通过该整合研究策略，可为土贝母作为辅助疗法抑制肝细胞癌发掘全新潜在治疗靶点，同时也有助于阐明土贝母治疗肝细胞癌的分子药理机制。