阐明复方苦参注射液对鼻咽癌 MYC-P15-CCND1 信号通路的药理作用——一项体外研究。

Elucidating the pharmacological effects of Compound Kushen injection on MYC-P15-CCND1 signaling pathway in nasopharyngeal carcinoma - An in vitro study.

机构信息

Department of Clinical Chinese Pharmacy, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100102, China.

Department of Medical Oncology, The Fourth Hospital of Hebei Medical University, Hebei Tumor Hospital, Shijiazhuang, China.

出版信息

J Ethnopharmacol. 2023 Oct 28;315:116702. doi: 10.1016/j.jep.2023.116702. Epub 2023 May 29.

DOI:10.1016/j.jep.2023.116702

PMID:37257705

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Compound Kushen injection (CKI) is a representative medication of Chinese herbal injection and is often used in the adjuvant treatment of nasopharyngeal carcinoma (NPC), but its antitumor mechanism is poorly understood.

AIM OF THE STUDY

To preliminarily elucidate the effects and possible mechanisms of CKI on NPC.

METHODS

In this work, we explored the possible molecular mechanisms of CKI against NPC by using network pharmacology and molecular docking. In addition, proteomics was used to explore the localization and quantitative information of protein in NPC C666-1 cells after the intervention of CKI, and enrichment analysis was used to obtain the potential targets and pathways. Finally, the effect and the core targets of CKI in the intervention of NPC were explored in vitro experiments.

RESULTS

Network pharmacology analysis identified three active components of CKI and 13 key targets. Molecular docking analysis showed that TNF, PTEN, CCND1, MAPK3, IL6, HIF1A, MYC had high affinity with corresponding components. Then the key pathway, cell cycle and the core targets MYC, CCND1, and P15 related to the key pathway were obtained. The results of in vitro experiments showed that CKI could inhibit the proliferation, migration, and invasion of NPC 5-8F cells and C666-1 cells, induce apoptosis of C666-1 cells, and arrest cell cycle G0/G1 phase. In addition, RT-qPCR and western blot showed that the expression of P15 was up-regulated and E2F4, E2F5, c-Myc, CCND1, and P107 was down-regulated in 5-8F cells and C666-1 cells intervened by CKI.

CONCLUSION

The key pathway, cell cycle and the corresponding core targets MYC, CCND1, and P15 were obtained from network pharmacology, molecular docking, and proteomics. CKI could inhibit the proliferation, migration, and invasion of NPC cells, induce apoptosis of C666-1 cells. Especially CKI may arrest cell cycle G0/G1 phase through regulating targets MYC/P15/CCND1 of cell cycle pathway.

摘要

民族药理学相关性

苦参注射液（CKI）是中药注射液的代表药物，常用于鼻咽癌（NPC）的辅助治疗，但对其抗肿瘤机制尚不清楚。

研究目的

初步阐明 CKI 对 NPC 的作用及可能机制。

方法

本研究采用网络药理学和分子对接方法探讨 CKI 对 NPC 的可能作用机制。此外，蛋白质组学用于探索 CKI 干预 NPC 后 C666-1 细胞中蛋白质的定位和定量信息，并进行富集分析以获得潜在的靶点和通路。最后，通过体外实验探讨 CKI 干预 NPC 的作用及核心靶点。

结果

网络药理学分析确定了 CKI 的三种活性成分和 13 个关键靶点。分子对接分析表明，TNF、PTEN、CCND1、MAPK3、IL6、HIF1A、MYC 与相应成分具有高亲和力。然后获得了关键通路、细胞周期以及与关键通路相关的核心靶点 MYC、CCND1 和 P15。体外实验结果表明，CKI 可抑制 NPC 5-8F 细胞和 C666-1 细胞的增殖、迁移和侵袭，诱导 C666-1 细胞凋亡，并阻滞细胞周期 G0/G1 期。此外，RT-qPCR 和 Western blot 结果显示，CKI 干预后 5-8F 细胞和 C666-1 细胞中 P15 表达上调，E2F4、E2F5、c-Myc、CCND1、P107 表达下调。