Oridonin impedes breast cancer growth by blocking cells in S phase and inhibiting the PI3K/AKT/mTOR signaling pathway

Weijie Zhang; Lei Shi; Wei Zhou; Xin Liu; Yuan Xi; Xinyin Wang; Ya Li; Xia Xu; Youcai Tang

doi:10.1016/j.heliyon.2023.e18046

Oridonin impedes breast cancer growth by blocking cells in S phase and inhibiting the PI3K/AKT/mTOR signaling pathway

Heliyon. 2023 Jul 11;9(7):e18046. doi: 10.1016/j.heliyon.2023.e18046. eCollection 2023 Jul.

Authors

Weijie Zhang¹, Lei Shi¹, Wei Zhou¹, Xin Liu², Yuan Xi¹, Xinyin Wang¹, Ya Li^{3

4

5

6

7}, Xia Xu⁸, Youcai Tang^{3

4

5

6

7}

Affiliations

¹ Department of Pharmacy, The Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, China.
² Department of Blood Transfusion, The Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, China.
³ Department of Pediatrics, The Fifth Affiliated Hospital, Zhengzhou University, 3 Kangfuqian Street, Zhengzhou, Henan 450052, China.
⁴ Henan Key Laboratory of Rehabilitation Medicine, The Fifth Affiliated Hospital, Zhengzhou University, 3 Kangfuqian Street, Zhengzhou, Henan 450052, China.
⁵ Henan Joint International Research Laboratory of Chronic Liver Injury, The Fifth Affiliated Hospital, Zhengzhou University, 3 Kangfuqian Street, Zhengzhou, Henan 450052, China.
⁶ Henan Workshop of Chronic Liver Injury for Outstanding Overseas Scientists, The Fifth Affiliated Hospital, Zhengzhou University, 3 Kangfuqian Street, Zhengzhou, Henan 450052, China.
⁷ Zhengzhou Key Laboratory of Metabolic-dysfunction-associated Fatty Liver Disease, The Fifth Affiliated Hospital, Zhengzhou University, 3 Kangfuqian Street, Zhengzhou, Henan 450052, China.
⁸ Key Laboratory of Advanced Drug Preparation Technologies, Ministry of Education of China, Co-innovation Center of Henan Province for New Drug R & D and Preclinical Safety, School of Pharmaceutical Sciences, Zhengzhou University, Zhengzhou, Henan 450001, China.

Abstract

Breast cancer is one of the most common cancers. Oridonin, a traditional Chinese medicine, is believed to inhibit tumor growth, but its particular effects on breast cancer remain unknown. In this study, we examined oridonin's effects on 4T1, MCF-7, and MDAMB-231 cellular activity using CCK8. Scratch assays were used to detect oridonin's effects on cellular migration. Oridonin's effects on the breast cancer cell cycle were studied using flow cytometry, and expression of cell cycle related proteins p53, CDK2, and p21 was detected using Western blot assays. Metabolomics assays were used to detect changes in small molecule metabolites and metabolic pathways in breast cancer cells after treatment with oridonin. Oridonin's effects on breast cancer growth were also studied using xenograft mice. Metabolomics assays were used to detect changes in metabolites and metabolic pathways in xenograft mouse plasma in a control group, model group, and drug administration group. Experimental results showed that oridonin could significantly inhibit breast cancer growth both in vivo and in vitro. Scratch experiments showed that oridonin could inhibit breast cancer cell migration. Oridonin was also able to arrest cells in S phase by affecting several cell cycle-related proteins, including p53, CDK2, and p21. Metabolomic analysis of 4T1 cells identified a total of 33 differential metabolites, including multiple amino acids (such as l-Glutamic acid, l-Asparagine, l-Histidine, l-Valine, and l-Isoleucine). KEGG pathway enrichment analysis showed significant changes in aminoacyl-tRNA biosynthesis, and in multiple amino acid metabolic pathways. Plasma metabolomic analyses of xenograft mice revealed 28 differentially-expressed metabolites between the different animal model groups, including multiple amino acids. KEGG pathway analysis showed significant alterations in multiple amino acid metabolic pathways in oridonin-treated mice. Additionally, changes in the expression of PI3K, AKT and mTOR proteins, as well as in branched amino acids, suggest that oridonin affects the PI3K/AKT/mTOR signaling pathway by inhibiting the biosynthesis of valine, leucine and isoleucine. Taken together, our results suggest that oridonin has strong anti-tumor activity in vitro and in vivo, and has potential as an adjuvant to breast cancer treatment regimens.

Keywords: Amino acid metabolism; Breast cancer; Metabolomics; Oridonin; PI3K/AKT/mTOR signaling pathway.