Uncovering the effects and molecular mechanism of Astragalus membranaceus (Fisch.) Bunge and its bioactive ingredients formononetin and calycosin against colon cancer: An integrated approach based on network pharmacology analysis coupled with experimental validation and molecular docking

Yu Hu; Wenjuan Zhai; Duanling Tan; Haipeng Chen; Guiyu Zhang; Xuanjing Tan; Yuting Zheng; Wenhui Gao; Yijie Wei; Jinjun Wu; Xin Yang

doi:10.3389/fphar.2023.1111912

Uncovering the effects and molecular mechanism of Astragalus membranaceus (Fisch.) Bunge and its bioactive ingredients formononetin and calycosin against colon cancer: An integrated approach based on network pharmacology analysis coupled with experimental validation and molecular docking

Front Pharmacol. 2023 Jan 23:14:1111912. doi: 10.3389/fphar.2023.1111912. eCollection 2023.

Authors

Yu Hu¹, Wenjuan Zhai¹, Duanling Tan², Haipeng Chen¹, Guiyu Zhang¹, Xuanjing Tan¹, Yuting Zheng¹, Wenhui Gao³, Yijie Wei¹, Jinjun Wu¹, Xin Yang²

Affiliations

¹ Guangdong Provincial Key Laboratory of Translational Cancer Research of Chinese Medicines, Joint International Research Laboratory of Translational Cancer Research of Chinese Medicines, School of Pharmaceutical Sciences, International Institute for Translational Chinese Medicine, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
² Key Laboratory of Molecular Target and Clinical Pharmacology and the State, NMPA Key Laboratory of Respiratory Disease, School of Pharmaceutical Sciences and The Fifth Affiliated Hospital, Guangzhou Medical University, Guangzhou, China.
³ Affiliated Cancer Hospital and Institute of Guangzhou Medical University, Guangzhou, Guangdong, China.

Abstract

Colon cancer is a highly malignant cancer with poor prognosis. Astragalus membranaceus (Fisch.) Bunge (Huang Qi in Chinese, HQ), a well-known Chinese herbal medicine and a popular food additive, possesses various biological functions and has been frequently used for clinical treatment of colon cancer. However, the underlying mechanism is not fully understood. Isoflavonoids, including formononetin (FMNT) and calycosin (CS), are the main bioactive ingredients isolated from HQ. Thus, this study aimed to explore the inhibitory effects and mechanism of HQ, FMNT and CS against colon cancer by using network pharmacology coupled with experimental validation and molecular docking. The network pharmacology analysis revealed that FMNT and CS exerted their anticarcinogenic actions against colon cancer by regulating multiple signaling molecules and pathways, including MAPK and PI3K-Akt signaling pathways. The experimental validation data showed that HQ, FMNT and CS significantly suppressed the viability and proliferation, and promoted the apoptosis in colon cancer Caco2 and HT-29 cells. HQ, FMNT and CS also markedly inhibited the migration of Caco2 and HT-29 cells, accompanied by a marked increase in E-cadherin expression, and a notable decrease in N-cadherin and Vimentin expression. In addition, HQ, FMNT and CS strikingly decreased the expression of ERK1/2 phosphorylation (p-ERK1/2) without marked change in total ERK1/2 expression. They also slightly downregulated the p-Akt expression without significant alteration in total Akt expression. Pearson correlation analysis showed a significant positive correlation between the inactivation of ERK1/2 signaling pathway and the HQ, FMNT and CS-induced suppression of colon cancer. The molecular docking results indicated that FMNT and CS had a strong binding affinity for the key molecules of ERK1/2 signaling pathway. Conclusively, HQ, FMNT and CS exerted good therapeutic effects against colon cancer by mainly inhibiting the ERK1/2 signaling pathway, suggesting that HQ, FMNT and CS could be useful supplements that may enhance chemotherapeutic outcomes and benefit colon cancer patients.

Keywords: Astragalus membranaceus (Fisch.) Bunge; ERK1/2; calycosin; colon cancer; formononetin; network pharmacology.

Grants and funding

This research was funded by the grants of Science and Technology Projects in Guangzhou (202201011393, 202102021196), Project of Traditional Chinese Medicine Bureau of Guangdong Province (20211111, 20231234, 20201201), Key-Area Research and Development Program of Guangdong Province (2020B1111100004), Guangdong Basic and Applied Basic Research Foundation (2020B1515130005), and First Class Undergraduate Major Construction Project of High Level University (2022JXA010).