目的 应用生物信息学与实验相结合的方法，分析常春藤皂苷元（hederagenin，HD）抗肝癌的作用机制。方法 通过UCSC Xena数据库获取肝癌RNA-seq数据，应用R软件“Limma”包筛选肝癌差异基因，SwissTargetPrediction数据库预测HD作用靶点，取二者交集基因，String数据库构建交集基因的蛋白质-蛋白质相互作用（protein-protein interaction，PPI）网络，Cytoscape_3.9.0软件筛选PPI网络中核心基因，采用分子对接方法验证HD与核心基因的结合位点。采用MTT法观察HD对人肝癌HepG2细胞增殖的影响；采用Western blotting法检测HD对HepG2细胞和H₂₂荷瘤小鼠模型瘤体内细胞分裂周期因子25A（cell division cycle 25A，CDC25A）、CDC25B、白细胞介素-6（interleukin-6，IL-6）、雄性激素受体（androgen receptor，AR）、雌激素受体1（estrogen receptor 1，ESR1）和前列腺素内过氧化物酶2（prostaglandin-endoperoxide synthase 2，PTGS2）蛋白表达的影响。结果 生物信息学分析显示，HD可能作用于IL-6、CDC25A、PTGS2、AR、CDC25B和ESR1，从而发挥抗肝癌作用。HD显著抑制HepG2细胞增殖（P＜0.05、0.01），下调HepG2细胞和瘤体IL-6、CDC25A、ESR1和PTGS2蛋白表达水平（P＜0.01），上调AR蛋白表达水平（P＜0.01），下调瘤体CDC25B蛋白表达水平（P＜0.01）。结论 HD能够通过调控IL-6、CDC25A、PTGS2、AR、CDC25B和ESR1，进而发挥抗肝癌的作用。

Objective To analyze the mechanism of anti-hepatocellular carcinoma effect of hederagenin based on bioinformatics combined with experiments. Methods UCSC Xena database was used to download RNA-seq data of hepatocellular carcinoma, R software “Limma” package was used to screen hepatocellular carcinoma differential genes. SwissTargetPrediction database was used to predict hederagenin target genes. Intersecting genes were screened between hepatocellular carcinoma differential genes and hederagenin target genes. String database was used to construct protein-protein interaction (PPI) network of intersecting genes, Cytoscape_3.9.0 was used to screen core genes in PPI network, and molecular docking was used to analyze the binding sites of hederagenin with core genes. MTT assay was used to detect the effect of hederagenin on proliferation of HepG2 cells; Western blotting was used to detect effect of hederagenin on cell division cycle 25A (CDC25A), CDC25B, interleukin-6 (IL-6), androgen receptor (AR), estrogen receptor 1 (ESR1) and prostaglandin-endoperoxide synthase 2 (PTGS2) protein expressions in HepG2 cells and tumor of H₂₂ tumor-bearing mice model. Results Bioinformatics analysis showed that hederagenin may act on IL-6, CDC25A, PTGS2, AR, CDC25B and ESR1 to treat hepatocellular carcinoma. Hederagenin significantly inhibited the proliferation of HepG2 cells (P < 0.05, 0.01), down-regulated IL-6, CDC25A, ESR1 and PTGS2 protein expression levels in HepG2 cells and tumors (P < 0.01), up-regulated AR protein expression level (P < 0.01), down-regulated CDC25B protein expression level in tumor (P < 0.01). Conclusion HD may exert anti-hepatocellular carcinogenic effect by regulating IL-6, CDC25A, PTGS2, AR, CDC25B and ESR1.

R285.5

黑龙江省自然科学基金资助项目（YQ2020H001）；黑龙江省教育科学“十三五”规划课题（GBD1317135）