Inhibition of Triple-negative Breast Cancer Growth Via Delphinidin-mediated Suppression of the JAK2/STAT3/PD-L1 Pathway

Overview

Journal Food Nutr Res

Date 2025 Jan 9

PMID 39781272

Authors

Xiaoping Yu

Xiaolong Song

Jiali Yan

Ziting Xiong

Lujie Zheng

Yan Luo

Fengcheng Deng

Yanfeng Zhu

Affiliations

Soon will be listed here.

Abstract

Background: Breast cancer is a leading cause of cancer-related mortality among women globally, with triple-negative breast cancer (TNBC) being particularly aggressive. Delphinidin (Dp), an anthocyanin monomer, has shown promising health benefits.

Objective: This study investigates the effects of Dp on TNBC and aims to elucidate its specific mechanisms of action.

Design: We utilized cell counting kit-8 (CCK-8) assays, colony formation assays, and scratch assays to evaluate the influence of Dp on the proliferation and migration of TNBC cells. Flow cytometry was employed to analyze programmed cell death-ligand 1 (PD-L1) and Cluster of Differentiation 69 expression, while Western blotting assessed the levels of PD-L1, Janus Kinase 2 (JAK2), Signal Transducer and Activator of Transcription 3 (STAT3), p-JAK2, p-STAT3, and exosomal marker proteins. Additionally, enzyme-linked immunosorbent assay (ELISA) was conducted to measure concentrations of PD-L1, interferon-γ (IFN-γ), and tumor necrosis factor-β (TNF-β).

Results: Dp effectively inhibited TNBC cell proliferation and migration, as evidenced by CCK-8, colony formation, and scratch assays. Flow cytometry and Western blot analysis indicated a reduction in PD-L1 expression in TNBC cells. Meanwhile, we successfully isolated TNBC cell-derived exosomes, with ELISA experiments showing a decrease in PD-L1 expression in these exosomes following Dp treatment. In a co-culture system with TNBC and Jurkat cells, Dp enhanced Cluster of Differentiation 69 expression and reactivated Jurkat cells, resulting in increased secretion of IFN-γ and TNF-β. Additionally, Dp significantly reduced the p-JAK2/JAK2 and p-STAT3/STAT3 ratios in TNBC cells.

Conclusion: Dp may exert its anti-TNBC effects by downregulating PD-L1 expression in TNBC cells and exosomes through the JAK2/STAT3 signaling pathway, potentially restoring T cell activity and modifying the tumor microenvironment.

References

Xie C, Zhou X, Liang C, Li X, Ge M, Chen Y . Correction to: Apatinib triggers autophagic and apoptotic cell death via VEGFR2/STAT3/PD-L1 and ROS/Nrf2/p62 signaling in lung cancer. J Exp Clin Cancer Res. 2021; 40(1):349. PMC: 8571894. DOI: 10.1186/s13046-021-02142-y. View

Mazewski C, Kim M, Gonzalez de Mejia E . Anthocyanins, delphinidin-3-O-glucoside and cyanidin-3-O-glucoside, inhibit immune checkpoints in human colorectal cancer cells in vitro and in silico. Sci Rep. 2019; 9(1):11560. PMC: 6689002. DOI: 10.1038/s41598-019-47903-0. View

Zhang N, Zeng Y, Du W, Zhu J, Shen D, Liu Z . The EGFR pathway is involved in the regulation of PD-L1 expression via the IL-6/JAK/STAT3 signaling pathway in EGFR-mutated non-small cell lung cancer. Int J Oncol. 2016; 49(4):1360-8. DOI: 10.3892/ijo.2016.3632. View

Wu J, Ge F, Zhu L, Liu N . Potential Toxic Mechanisms of Neonicotinoid Insecticides in Rice: Inhibiting Auxin-Mediated Signal Transduction. Environ Sci Technol. 2023; 57(12):4852-4862. DOI: 10.1021/acs.est.2c09352. View

He C, Zheng S, Luo Y, Wang B . Exosome Theranostics: Biology and Translational Medicine. Theranostics. 2018; 8(1):237-255. PMC: 5743472. DOI: 10.7150/thno.21945. View

Wen J, Chang X, Bai B, Gao Q, Zhao Y . Orexin A Suppresses the Expression of Exosomal PD-L1 in Colon Cancer and Promotes T Cell Activity by Inhibiting JAK2/STAT3 Signaling Pathway. Dig Dis Sci. 2021; 67(6):2173-2181. DOI: 10.1007/s10620-021-07077-0. View

Liu X, Wang J, Wang H, Yin G, Liu Y, Lei X . REG3A accelerates pancreatic cancer cell growth under IL-6-associated inflammatory condition: Involvement of a REG3A-JAK2/STAT3 positive feedback loop. Cancer Lett. 2015; 362(1):45-60. DOI: 10.1016/j.canlet.2015.03.014. View

Xu L, Chen X, Shen M, Yang D, Fang L, Weng G . Inhibition of IL-6-JAK/Stat3 signaling in castration-resistant prostate cancer cells enhances the NK cell-mediated cytotoxicity via alteration of PD-L1/NKG2D ligand levels. Mol Oncol. 2017; 12(3):269-286. PMC: 5830627. DOI: 10.1002/1878-0261.12135. View

Zagorski J, Turley A, Dover H, VanDenBerg K, Compton J, Rockwell C . The Nrf2 activator, tBHQ, differentially affects early events following stimulation of Jurkat cells. Toxicol Sci. 2013; 136(1):63-71. PMC: 3829568. DOI: 10.1093/toxsci/kft172. View

10.

Rak J . Extracellular vesicles - biomarkers and effectors of the cellular interactome in cancer. Front Pharmacol. 2013; 4:21. PMC: 3589665. DOI: 10.3389/fphar.2013.00021. View

11.

Ozbay T, Nahta R . Delphinidin Inhibits HER2 and Erk1/2 Signaling and Suppresses Growth of HER2-Overexpressing and Triple Negative Breast Cancer Cell Lines. Breast Cancer (Auckl). 2011; 5:143-54. PMC: 3140266. DOI: 10.4137/BCBCR.S7156. View

12.

Kudelova E, Smolar M, Holubekova V, Hornakova A, Dvorska D, Lucansky V . Genetic Heterogeneity, Tumor Microenvironment and Immunotherapy in Triple-Negative Breast Cancer. Int J Mol Sci. 2022; 23(23). PMC: 9735793. DOI: 10.3390/ijms232314937. View

13.

Gholijani N, Gharagozloo M, Kalantar F, Ramezani A, Amirghofran Z . Modulation of Cytokine Production and Transcription Factors Activities in Human Jurkat T Cells by Thymol and Carvacrol. Adv Pharm Bull. 2016; 5(Suppl 1):653-60. PMC: 4708037. DOI: 10.15171/apb.2015.089. View

14.

Li C, Li C, Zhi C, Liang W, Wang X, Chen X . Clinical significance of PD-L1 expression in serum-derived exosomes in NSCLC patients. J Transl Med. 2019; 17(1):355. PMC: 6820965. DOI: 10.1186/s12967-019-2101-2. View

15.

Niu M, Liu Y, Yi M, Jiao D, Wu K . Biological Characteristics and Clinical Significance of Soluble PD-1/PD-L1 and Exosomal PD-L1 in Cancer. Front Immunol. 2022; 13:827921. PMC: 8977417. DOI: 10.3389/fimmu.2022.827921. View

16.

Zhang Z, Pan Y, Zhao Y, Ren M, Li Y, Lu G . Delphinidin modulates JAK/STAT3 and MAPKinase signaling to induce apoptosis in HCT116 cells. Environ Toxicol. 2021; 36(8):1557-1566. DOI: 10.1002/tox.23152. View

17.

Koss-Mikolajczyk I, Bartoszek A . Relationship between Chemical Structure and Biological Activity Evaluated In Vitro for Six Anthocyanidins Most Commonly Occurring in Edible Plants. Molecules. 2023; 28(16). PMC: 10458735. DOI: 10.3390/molecules28166156. View

18.

Mahoney K, Freeman G, Mcdermott D . The Next Immune-Checkpoint Inhibitors: PD-1/PD-L1 Blockade in Melanoma. Clin Ther. 2015; 37(4):764-82. PMC: 4497957. DOI: 10.1016/j.clinthera.2015.02.018. View

19.

Sung H, Ferlay J, Siegel R, Laversanne M, Soerjomataram I, Jemal A . Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021; 71(3):209-249. DOI: 10.3322/caac.21660. View

20.

Hu X, Li J, Fu M, Zhao X, Wang W . The JAK/STAT signaling pathway: from bench to clinic. Signal Transduct Target Ther. 2021; 6(1):402. PMC: 8617206. DOI: 10.1038/s41392-021-00791-1. View