Fu Chunqing, Zhang Keyu, Wang Manyuan, Qiu Feng
Beijing Key Lab of TCM Collateral Disease Theory Research, School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China.
Beijing Key Lab of TCM Collateral Disease Theory Research, School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China.
Phytomedicine. 2022 Jun;100:154095. doi: 10.1016/j.phymed.2022.154095. Epub 2022 Apr 4.
Artemisia annua L. (A. annua) and its active components exhibit antitumour effects in many cancer cells. However, the biological processes and mechanisms involved are not well understood, especially for the treatment of non-small-cell lung cancer (NSCLC).
This study aimed to comprehensively explore the biological processes of A. annua and its active components in NSCLC cells and to identify the mechanism by which these compounds induce apoptosis.
STUDY DESIGNS/METHODS: Cell viability and flow cytometry assays were used to evaluate the cytotoxicity of A. annua active components casticin (CAS) and chrysosplenol D (CHD) in A. annua in NSCLC cells. After treatment with CAS and CHD, A549 cells were subjected to RNA sequencing (RNA-seq) analysis, differentially expressed genes (DEGs) were screened and subjected to functional enrichment analysis (KEGG and GO analysis) as well as protein interaction network analysis. The key targets associated with apoptosis induction in A549 cells were screened by Cytoscape, and the screened DEGs were validated by qRT-PCR. Immunoblotting, immunofluorescence, and molecular docking assays were used to determine whether CAS and/or CHD could induce apoptosis in NSCLC cells by inducing DNA damage through down-regulation of topoisomerase IIα (topo IIα) expression. The same experiments were verified again in the H1299 lung cancer cell line.
CAS and CHD inhibited NSCLC cells proliferation in a time- and dose-dependent manner, and significantly induced apoptosis. A total of 115 co-upregulated DEGs and 277 co-downregulated DEGs were identified in A549 cells following treatment with CAS and CHD. Comprehensive and systematic data about biological processes and mechanisms were obtained. DNA damage pathways and topo IIα targets were screened to study the apoptosis effects of CAS and CHD on NSCLC cells. CAS and CHD may be able to induce DNA damage by binding to topo IIα-DNA and reducing topo IIα activity.
This study suggested that CAS and CHD may reduce topo IIα activity by binding to topo IIα-DNA, affecting the replication of DNA, triggering DNA damage, and inducing apoptosis. It described a novel mechanism associated with topo IIα inhibition to reveal a novel role for CAS and CHD in A. annua as potential anticancer agents and/or adjuvants in NSCLC cells.
黄花蒿及其活性成分在多种癌细胞中表现出抗肿瘤作用。然而，其中涉及的生物学过程和机制尚未完全明确，尤其是在非小细胞肺癌（NSCLC）的治疗方面。
本研究旨在全面探究黄花蒿及其活性成分在NSCLC细胞中的生物学过程，并确定这些化合物诱导细胞凋亡的机制。
研究设计/方法：采用细胞活力测定和流式细胞术分析来评估黄花蒿活性成分紫花牡荆素（CAS）和金腰乙素（CHD）对NSCLC细胞的细胞毒性。用CAS和CHD处理后，对A549细胞进行RNA测序（RNA-seq）分析，筛选差异表达基因（DEGs）并进行功能富集分析（KEGG和GO分析）以及蛋白质相互作用网络分析。通过Cytoscape筛选与A549细胞凋亡诱导相关的关键靶点，并用qRT-PCR验证筛选出的DEGs。采用免疫印迹、免疫荧光和分子对接试验来确定CAS和/或CHD是否可通过下调拓扑异构酶IIα（topo IIα）表达诱导DNA损伤，从而诱导NSCLC细胞凋亡。在H1299肺癌细胞系中再次验证相同的实验。
CAS和CHD以时间和剂量依赖性方式抑制NSCLC细胞增殖，并显著诱导细胞凋亡。在用CAS和CHD处理后的A549细胞中，共鉴定出115个共同上调的DEGs和277个共同下调的DEGs。获得了关于生物学过程和机制的全面系统的数据。筛选了DNA损伤途径和topo IIα靶点以研究CAS和CHD对NSCLC细胞的凋亡作用。CAS和CHD可能能够通过与topo IIα-DNA结合并降低topo IIα活性来诱导DNA损伤。
本研究表明，CAS和CHD可能通过与topo IIα-DNA结合降低topo IIα活性，影响DNA复制，引发DNA损伤并诱导细胞凋亡。它描述了一种与topo IIα抑制相关的新机制，揭示了黄花蒿中的CAS和CHD作为NSCLC细胞中潜在抗癌药物和/或佐剂的新作用。
