Molecular mechanism of Puerariae Lobatae Radix in treatment of hepatocellular carcinoma based on network pharmacology

ZHOU Min1,2 ZHOU Yuan3 CHENG Ye2 XU Nan2 LI Wei-bing2 WU Cheng-yu1 CHEN Yan2

(1.College of Traditional Chinese Medicine·College of Integrated Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China 210046)
(2.Jiangsu Cancer Hospital & Jiangsu Institute of Cancer Research & the Affiliated Cancer Hospital of Nanjing Medical University, Nanjing, Jiangsu, China 210009)
(3.Department of General Surgery, the Affiliated Sir Run Run Shaw Hospital of School of Medicine of Zhejiang University, Hangzhou, Zhejiang, China 310016)

【Abstract】To investigate the potential mechanism of Puerariae Lobatae Radix in the treatment of hepatocellular carcinoma (HCC) by network pharmacology and in vitro cell experiment. The main active components of Puerariae Lobatae Radix and their predicted targets were obtained from TCMSP, and the disease targets were obtained from GeneCards database. The disease and drug prediction targets were intersected to select the common potential therapeutic targets. The “compound-target-disease” network diagram was constructed in Cytoscape 3.7.1, and the common targets were input into the STRING database to build the PPI network. GO function and KEGG pathway enrichment analysis on effective targets were performed by R software. Autodock vina 1.1.2 was used for molecular docking. Finally, the core targets and pathways were preliminarily verified by in vitro experiments. The proliferation of human HCC cells was detected by CCK-8 and EdU enzyme staining, and the expressions of PTEN, PDK1, Akt and GSK3 were detected by Western blot. In this study, 10 components of Puerariae Lobatae Radix (9 components involved in HCC-related targets and signaling pathways), 149 HCC-related targets, and 156 signaling pathways were screened out. The results of network analysis indicated that Puerariae Lobatae Radix may play an anti-HCC effect on key targets, such as Akt, IL6, MAPK3, EGFR, and key pathways, such as PI3K-Akt. The results of molecular docking indicated that puerarin, genistein and daidzein had good binding abilities with the key targets such as AKT1, MAPK3, MAPK1 and CASP3, and puerarin had the lowest Vina score with AKT1 and MAPK3 and also similar to them. In vitro cell experiments confirmed that puerarin had a significantly inhibitory effect on the proliferation of human HCC cells. Western blot results showed that puerarin could increase the phosphorylation of PTEN in human HCC cells through the PTEN/Akt/GSK3β signaling pathway, and the phosphorylation level of its downstream Akt decreased. This series of studies confirm that puerarin can treat HCC by blocking PTEN/Akt/GSK3β cellular signaling pathway, so as to provide ideas for subsequent studies for the molecular mechanism of puerarin in the treatment of liver cancer.

【Keywords】 network pharmacology; Puerariae Lobatae Radix; puerarin; hepatocellular carcinoma; in vitro cell experiment;

【DOI】

【Funds】 General Project of the National Natural Science Foundation of China (81874452) General Project of Jiangsu Cancer Hospital (ZM201810)

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This Article

ISSN:1001-5302

CN: 11-2272/R

Vol 45, No. 17, Pages 4089-4098

September 2020

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Article Outline

Abstract

  • 1 Materials and methods
  • 2 Results
  • 3 Discussion
  • References