School of Basic medical, Inner Mongolia Medical University, Hohhot, Inner Mongolia, China.
Inner Mongolia Hospital of Traditional Chinese Medicine, Hohhot, Inner Mongolia, China.
BMC Complement Med Ther. 2023 May 18;23(1):160. doi: 10.1186/s12906-023-03975-0.
Cantharidin (CTD) is a major ingredient of cantharis (Mylabris phalerata Pallas) and has been used extensively in traditional Chinese medicines. It has been shown to exhibit anticancer activity in multiple types of cancer, especially hepatocellular carcinoma (HCC). However, there is no systematic study on the relationships among the regulatory networks of its targets in HCC therapy. We focused on histone epigenetic regulation and the influence of CTD on the immune response in HCC.
We performed a comprehensive analysis of novel CTD targets in HCC based on network pharmacology and RNA-seq approaches. The mRNA levels of target genes were analyzed by qRT-PCR, and the corresponding protein levels were confirmed using enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining (IHC). ChIP-seq data were visualized by IGV software. The associations of gene transcript levels with the cancer immune score and infiltration level were investigated using TIMER. In vivo, the H22 mouse model of hepatocellular carcinoma was established by treatment with CTD and 5-Fu. The immune cell proportions in the blood were elevated in model mice, as shown by flow cytometry.
We identified 58 targets of CTD, which were involved in various pathways in cancer, including apoptosis, the cell cycle, EMT and immune pathways. Moreover, we found that 100 EMT-related genes were differentially expressed after CTD treatment in HCC cells. Interestingly, our results confirmed that the EZH2/H3K27me3 -related cell cycle pathway is a therapeutic target of CTD in antitumour. In addition, we evaluated the influence of CTD on the immune response. Our data showed that the significantly enriched gene sets were positively correlated with the chemokine biosynthetic and chemokine metabolic modules. The proportions of CD4+/CD8 + T cells and B cells were increased, but the proportion of Tregs was decreased after treatment with CTD in vivo. Moreover, we found that the expression of the inflammatory factor and immune checkpoint genes PD-1/PD-L1 was significantly reduced in the mouse model.
We performed a novel integrated analysis of the potential role of CTD in HCC treatment. Our results provide innovative insight into the mechanism by which cantharidin exerts antitumour effects by regulating target genes expression to mediate apoptosis, EMT, cell cycle progression and the immune response in HCC. Based on the effect of CTD on the immune response, it can be used as a potential effective drug to activate antitumour immunity for the treatment of liver cancer.
斑蝥素（CTD）是芫青（Mylabris phalerata Pallas）的主要成分，广泛应用于传统中药。已证实其在多种类型的癌症中具有抗癌活性，特别是肝癌（HCC）。然而，目前尚无关于 HCC 治疗中其靶点调控网络之间关系的系统研究。我们专注于组蛋白表观遗传调控以及 CTD 对 HCC 免疫反应的影响。
我们基于网络药理学和 RNA-seq 方法对 HCC 中新型 CTD 靶点进行了全面分析。通过 qRT-PCR 分析靶基因的 mRNA 水平，并使用酶联免疫吸附测定（ELISA）和免疫组织化学染色（IHC）确认相应的蛋白水平。使用 IGV 软件可视化 ChIP-seq 数据。使用 TIMER 研究基因转录水平与癌症免疫评分和浸润水平的相关性。在体内，通过 CTD 和 5-Fu 处理建立 H22 肝癌小鼠模型。通过流式细胞术显示模型小鼠血液中的免疫细胞比例升高。
我们鉴定了 58 个 CTD 靶点，这些靶点参与了癌症中的各种途径，包括细胞凋亡、细胞周期、EMT 和免疫途径。此外，我们发现 CTD 处理 HCC 细胞后，有 100 个 EMT 相关基因表达差异。有趣的是，我们的结果证实 EZH2/H3K27me3 相关细胞周期途径是 CTD 在抗肿瘤中的治疗靶点。此外，我们评估了 CTD 对免疫反应的影响。我们的数据表明，显著富集的基因集与趋化因子生物合成和趋化因子代谢模块呈正相关。在体内用 CTD 处理后，CD4+/CD8+T 细胞和 B 细胞的比例增加，但 Treg 细胞的比例减少。此外，我们发现炎性因子和免疫检查点基因 PD-1/PD-L1 的表达在小鼠模型中显著降低。
我们对 CTD 在 HCC 治疗中的潜在作用进行了新颖的综合分析。我们的结果为斑蝥素通过调节靶基因表达来介导 HCC 中的细胞凋亡、EMT、细胞周期进程和免疫反应提供了创新的见解。基于 CTD 对免疫反应的影响，它可以用作激活抗肿瘤免疫以治疗肝癌的潜在有效药物。
