Yu Xiaoping, Song Xiaolong, Yan Jiali, Xiong Ziting, Zheng Lujie, Luo Yan, Deng Fengcheng, Zhu Yanfeng
School of Medicine and Nursing, Chengdu University, Chengdu, China.
School of Public Health, Chengdu Medical College, Chengdu, China.
Food Nutr Res. 2024 Dec 31;68. doi: 10.29219/fnr.v68.10974. eCollection 2024.
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.
This study investigates the effects of Dp on TNBC and aims to elucidate its specific mechanisms of action.
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).
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.
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.
乳腺癌是全球女性癌症相关死亡的主要原因，三阴性乳腺癌（TNBC）尤其具有侵袭性。飞燕草素（Dp）是一种花青素单体，已显示出有益健康的作用。
本研究调查Dp对TNBC的影响，并旨在阐明其具体作用机制。
我们使用细胞计数试剂盒-8（CCK-8）检测、集落形成检测和划痕检测来评估Dp对TNBC细胞增殖和迁移的影响。采用流式细胞术分析程序性细胞死亡配体1（PD-L1）和分化簇69的表达，同时蛋白质印迹法评估PD-L1、Janus激酶2（JAK2）、信号转导子和转录激活子3（STAT3）、磷酸化JAK2、磷酸化STAT3和外泌体标记蛋白的水平。此外，进行酶联免疫吸附测定（ELISA）以测量PD-L1、干扰素-γ（IFN-γ）和肿瘤坏死因子-β（TNF-β）的浓度。
CCK-8检测、集落形成检测和划痕检测表明，Dp有效抑制TNBC细胞增殖和迁移。流式细胞术和蛋白质印迹分析表明TNBC细胞中PD-L1表达降低。同时，我们成功分离出TNBC细胞来源的外泌体，ELISA实验显示Dp处理后这些外泌体中PD-L1表达降低。在TNBC与Jurkat细胞的共培养系统中，Dp增强了分化簇69的表达并重新激活Jurkat细胞，导致IFN-γ和TNF-β分泌增加。此外，Dp显著降低TNBC细胞中磷酸化JAK2/JAK2和磷酸化STAT3/STAT3的比值。
Dp可能通过JAK2/STAT3信号通路下调TNBC细胞和外泌体中PD-L1的表达来发挥其抗TNBC作用，可能恢复T细胞活性并改变肿瘤微环境。
