氯喹通过PI3K/AKT/MDM2信号通路对宫颈癌的影响。
The effect of chloroquine on cervical cancer via the PI3K/AKT/MDM2 pathway.
作者信息
Xu Dandan, Hou Chenyang, Chang Jianliang, Yang Shuaibo, Yang Zhicong, Yuan Zelong, Li Jingqi, Liu Shuqin, Liu Yaqun, Su Yingpeng, Gao Peng, Liu Xiaojuan
机构信息
Central Laboratory, The First Affiliated Hospital of Hebei North University, Zhangjiakou City, 075000, Hebei Province, China.
Hebei Key Laboratory of Systems Biology and Gene Regulation, The First Affiliated Hospital of Hebei North University, zhangjiakou, China.
出版信息
Discov Oncol. 2025 Jun 23;16(1):1179. doi: 10.1007/s12672-025-02939-9.
BACKGROUND
Cervical cancer (CC), the most prevalent malignant tumor in the female reproductive system, is characterized by its high incidence and mortality rates. Chloroquine(CQ), a 4-aminoquinoline drug originally used to prevent and treat malaria, has gained recognition as an adjuvant anti-cancer agent. However, whether chloroquine exhibits anti-CC activity and its underlying mechanisms remain unclear. This study aimed to elucidate chloroquine's anti-cancer effects, focusing on the PI3K/AKT/MDM2 pathway, by integrating network pharmacology and in vitro experiments.
METHODS
Firstly, CQ and its associated targets related to cervical cancer were identified using the PharmMapper, SwissTargetPrediction, DrugBank, GeneCards, and DisGeNET databases. A protein-protein interaction (PPI) network was constructed to screen for key targets. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted to predict the underlying biological functions and mechanisms involved in the anti-colorectal cancer effects of CQ. Subsequently, CCK-8, colony formation, and wound-healing assays were performed to assess the impact of CQ treatment on the proliferation ability of HeLa cells. Cell apoptosis was analyzed using flow cytometry, and the expression of apoptosis-related proteins was determined through Western blotting. Finally, the effects of CQ on the PI3K/AKT/MDM2 signaling pathway were evaluated via Western blotting.
RESULTS
A total of 7,846 potential treatment targets for CQ were identified, among which 126 were related to CC. Through PPI network analysis of 100 common targets, eight core targets were identified: TNF, MDM2, SLC6A4, GRIN1, CHRM1, EZH2, ACHE, and ADRB2. KEGG enrichment analysis revealed the PI3K/AKT signaling pathway as a central pathway of interest. Various concentrations of CQ (25, 50, and 75 µM) inhibited the viability of HeLa cells while showing no significant effect on H8 cells. Following treatment with CQ (25 µM and 75 µM), a reduction in proliferation and colony formation was observed in HeLa cells. Furthermore, CQ treatment led to an increase in the expression of apoptosis-related proteins (Bax, Bcl-2, cleaved-PARP) and a decrease in the expression of proteins associated with the PI3K/AKT/MDM2 signaling pathway (p-PI3K, p-AKT, and p-MDM2).
CONCLUSIONS
This study suggests that chloroquine may induce apoptosis by inhibiting the PI3K/AKT/MDM2 signalling pathway.
背景
宫颈癌（CC）是女性生殖系统中最常见的恶性肿瘤，具有高发病率和高死亡率的特点。氯喹（CQ）是一种最初用于预防和治疗疟疾的4-氨基喹啉类药物，已被认可为一种辅助抗癌药物。然而，氯喹是否具有抗CC活性及其潜在机制仍不清楚。本研究旨在通过整合网络药理学和体外实验，阐明氯喹的抗癌作用，重点关注PI3K/AKT/MDM2信号通路。
方法
首先，使用PharmMapper、SwissTargetPrediction、DrugBank、GeneCards和DisGeNET数据库鉴定CQ及其与宫颈癌相关的靶点。构建蛋白质-蛋白质相互作用（PPI）网络以筛选关键靶点。此外，进行基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析，以预测CQ抗结直肠癌作用所涉及的潜在生物学功能和机制。随后，进行CCK-8、集落形成和伤口愈合实验，以评估CQ处理对HeLa细胞增殖能力的影响。使用流式细胞术分析细胞凋亡，并通过蛋白质免疫印迹法测定凋亡相关蛋白的表达。最后，通过蛋白质免疫印迹法评估CQ对PI3K/AKT/MDM2信号通路的影响。
结果
共鉴定出7846个CQ的潜在治疗靶点，其中126个与CC相关。通过对100个共同靶点的PPI网络分析，确定了8个核心靶点：TNF、MDM2、SLC6A4、GRIN1、CHRM1、EZH2、ACHE和ADRB2。KEGG富集分析显示PI3K/AKT信号通路是一个重要的核心通路。不同浓度的CQ（25、50和75μM）抑制了HeLa细胞的活力，而对H8细胞无显著影响。用CQ（25μM和75μM）处理后，观察到HeLa细胞的增殖和集落形成减少。此外，CQ处理导致凋亡相关蛋白（Bax、Bcl-2、裂解的PARP）表达增加，与PI3K/AKT/MDM2信号通路相关的蛋白（p-PI3K、p-AKT和p-MDM2）表达减少。
结论
本研究表明，氯喹可能通过抑制PI3K/AKT/MDM2信号通路诱导细胞凋亡。
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