清胰化积方通过抑制 MAPK/ERK 和 PI3K/Akt/mTOR 信号通路促进体内外胰腺癌的细胞凋亡和自噬。

Qingyihuaji Formula promotes apoptosis and autophagy through inhibition of MAPK/ERK and PI3K/Akt/mTOR signaling pathway on pancreatic cancer in vivo and in vitro.

机构信息

Zhejiang Chinese Medical University, Zhejiang, China.

The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Zhejiang, China.

出版信息

J Ethnopharmacol. 2023 May 10;307:116198. doi: 10.1016/j.jep.2023.116198. Epub 2023 Jan 21.

DOI:10.1016/j.jep.2023.116198

PMID:36690307

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Qingyihuaji Formula (QYHJ), a widely used traditional Chinese medicine (TCM), has been used to treat patients with cancer in China. However, the effect and mechanism of QYHJ on pancreatic ductal adenocarcinoma (PDAC) remains unclear.

AIM OF THE STUDY

This study aimed to explore the roles and evaluate the possible underlying molecular mechanisms of QYHJ and its core component in PDAC using label-free quantitative proteomics in conjunction with network pharmacology-based analysis.

MATERIALS AND METHODS

By screening differentially expressed proteins (DEPs) in proteomics and QYHJ-predicted gene sets, we identified QYHJ-related PDAC targets annotated with bioinformatic analysis. A subcutaneous tumor model was established to assess the role of QYHJ in vivo. The effects of quercetin (Que), a core component of QYHJ, on cell proliferation, migration, invasion, apoptosis, and autophagy in SW1990 and PANC-1 cells were investigated in vitro. Immunohistochemistry, western blotting, mRFP-GFP-LC3 adenovirus, and kinase analysis were used to determine the underlying mechanisms.

RESULTS

Bioinformatics analysis revealed that 41 QYHJ-related PDAC targets were closely related to the cellular response to nitrogen compounds, positive regulation of cell death, regulation of epithelial cell apoptotic processes, and chemokine signaling pathways. CASP3, SRC, STAT1, PTPN11, PKM, and PAK1 with high expression were identified as hub DEPs in the PPI network, and these DEPs were associated with poor overall survival and STAT 1, MAPK/ERK, and PI3K/Akt/mTOR signaling pathways in PDAC patients. QYHJ significantly promoted tumor death in nude mice. Moreover, quercetin inhibited the proliferation, migration, and invasion of PDAC cells. Additionally, Que induced apoptosis and autophagy in PDAC cells. Mechanistically, QYHJ and Que significantly activated STAT 1 and remarkably inhibited the MAPK/ERK and PI3K/Akt/mTOR signaling pathways in vivo and in vitro, respectively. Importantly, ERK1/2 inactivation contributes to que-induced apoptosis in SW1990 and PANC-1 cells.

CONCLUSIONS

These results suggest that QYHJ and Que are promising anti-PDAC avenues that benefit from their multiform mechanisms.

摘要

民族药理学相关性

青银化瘀方（QYHJ）是一种广泛应用于中国癌症患者的中药。然而，QYHJ 对胰腺导管腺癌（PDAC）的作用和机制尚不清楚。

研究目的

本研究旨在通过无标记定量蛋白质组学结合网络药理学分析，探讨 QYHJ 及其核心成分在 PDAC 中的作用，并评估其可能的潜在分子机制。

材料与方法

通过蛋白质组学筛选差异表达蛋白（DEPs）和 QYHJ 预测基因集，我们鉴定了生物信息学分析注释的与 QYHJ 相关的 PDAC 靶标。建立皮下肿瘤模型评估 QYHJ 在体内的作用。体外研究槲皮素（Que），QYHJ 的核心成分，对 SW1990 和 PANC-1 细胞增殖、迁移、侵袭、凋亡和自噬的影响。免疫组织化学、Western blot、mRFP-GFP-LC3 腺病毒和激酶分析用于确定潜在机制。

结果

生物信息学分析表明，41 个 QYHJ 相关的 PDAC 靶标与细胞对含氮化合物的反应、细胞死亡的正调控、上皮细胞凋亡过程的调控以及趋化因子信号通路密切相关。在 PPI 网络中，CASP3、SRC、STAT1、PTPN11、PKM 和 PAK1 等高表达被鉴定为关键 DEPs，这些 DEPs与 PDAC 患者的总生存期不良和 STAT1、MAPK/ERK 和 PI3K/Akt/mTOR 信号通路相关。QYHJ 显著促进裸鼠肿瘤死亡。此外，槲皮素抑制 PDAC 细胞的增殖、迁移和侵袭。此外，Que 诱导 PDAC 细胞凋亡和自噬。机制上，QYHJ 和 Que 分别在体内和体外显著激活 STAT1，并显著抑制 MAPK/ERK 和 PI3K/Akt/mTOR 信号通路。重要的是，ERK1/2 失活有助于 Que 诱导 SW1990 和 PANC-1 细胞凋亡。