Institute of Integrated Medicine, Qingdao University, Qingdao, 266071, China.
Department of Oncology, The Affiliated Qingdao Hiser hospital of Qingdao University (Qingdao Hospital of Traditional Chinese Medicine), Qingdao, 266071, China.
Sci Rep. 2023 Jul 31;13(1):12351. doi: 10.1038/s41598-023-39655-9.
Triple-negative breast cancer (TNBC), a highly aggressive and heterogeneous subtype of breast cancer, lacks effective treatment options. Sophora flavescens Aiton, a Chinese medicinal plant, is often used in traditional Chinese medicine to treat cancer. Matrine (MAT) is an alkaloid extracted from Sophora flavescens. It has good anticancer effects, and thus can be explored as a new therapeutic agent in TNBC research. We performed bioinformatics analysis to analyze the differentially expressed genes between normal breast tissues and TNBC tissues, and comprehensive network pharmacology analyses. The activity and invasion ability of TNBC cells treated with MAT were analyzed. Apoptosis and cell cycle progression were determined using cytometry. We used Monodansylcadaverine (MDC) staining to determine the condition of autophagosomes. Finally, the expression levels of the key target proteins of the PI3K/AKT pathway were determined using western blotting. The proliferation and invasion ability of MDA-MB-231 and MDA-MB-468 can be effectively inhibited by MAT. The results of flow cytometry indicated that MAT arrested the TNBC cell cycle and induced apoptosis. In addition, we confirmed that MAT inhibited the expression of BCL-2 while up-regulating the expression of cleaved caspase-3. Moreover, enhanced intensity of MDC staining and high LC3-II expression were observed, which confirmed that MAT induced autophagy in TNBC cells. Western blotting showed that MAT inhibited the PI3K/AKT pathway and downregulated the expressions of PI3K, AKT, p-AKT, and PGK1. This study provides feasible methods, which include bioinformatics analysis and in vitro experiments, for the identification of compounds with anti-TNBC properties. MAT inhibited the PI3K/AKT signaling pathway, arrested cell cycle, as well as promoted cell apoptosis and autophagy. These experiments provide evidence for the anti-TNBC effect of MAT and identified potential targets against TNBC.
三阴性乳腺癌（TNBC）是一种侵袭性强、异质性高的乳腺癌亚型，缺乏有效的治疗方法。苦参是一种常用于中药治疗癌症的中国药用植物。苦参碱（MAT）是从苦参中提取的一种生物碱。它具有良好的抗癌作用，因此可以作为 TNBC 研究中的一种新的治疗剂进行探索。我们进行了生物信息学分析，以分析正常乳腺组织和 TNBC 组织之间的差异表达基因，并进行了综合网络药理学分析。分析了 MAT 处理的 TNBC 细胞的活性和侵袭能力。通过细胞术测定细胞凋亡和细胞周期进程。使用 Monodansylcadaverine（MDC）染色来确定自噬体的状态。最后，使用 Western blot 测定 PI3K/AKT 通路的关键靶蛋白的表达水平。MAT 可以有效抑制 MDA-MB-231 和 MDA-MB-468 的增殖和侵袭能力。流式细胞术结果表明，MAT 使 TNBC 细胞周期停滞并诱导细胞凋亡。此外，我们证实 MAT 抑制了 BCL-2 的表达，同时上调了 cleaved caspase-3 的表达。此外，观察到 MDC 染色强度增强和 LC3-II 表达增加，这证实了 MAT 在 TNBC 细胞中诱导了自噬。Western blot 显示 MAT 抑制了 PI3K/AKT 通路，并下调了 PI3K、AKT、p-AKT 和 PGK1 的表达。这项研究为鉴定具有抗 TNBC 特性的化合物提供了可行的方法，包括生物信息学分析和体外实验。MAT 抑制了 PI3K/AKT 信号通路，使细胞周期停滞，并促进细胞凋亡和自噬。这些实验为 MAT 的抗 TNBC 作用提供了证据，并确定了针对 TNBC 的潜在靶标。
