Benzo(a)pyrene promotes malignant progression of human lung cancer LTEP-a-2 cells through the NLRP3 pyroptosis signaling pathway: Validation based on bioinformatics and cellular experiments

DENG Shuangwu; GUO Fei; PENG Wantao; TAN Yupei; LIU Qingqing; ZHONG Jiangxue; YAO Maolin; ZHOU Ziyuan; CAO Jia; CHEN Xiong; LIU Wenbin

doi:10.16016/j.2097-0927.202511095

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Benzo(a)pyrene promotes malignant progression of human lung cancer LTEP-a-2 cells through the NLRP3 pyroptosis signaling pathway: Validation based on bioinformatics and cellular experiments

Basic Medicine | 更新时间：2026-03-30

- Benzo(a)pyrene promotes malignant progression of human lung cancer LTEP-a-2 cells through the NLRP3 pyroptosis signaling pathway: Validation based on bioinformatics and cellular experiments
- Journal of Army Medical University Vol. 48, Issue 6, Pages: 709-721(2026)
- 作者机构：
  
  1.贵州医科大学公共卫生与健康学院，环境污染与疾病监控教育部重点实验室，贵州贵阳
  2.陆军军医大学（第三军医大学）军事预防医学系军队环境卫生学教研室，重庆
  3.重庆理工大学药学与生物工程学院药学系，重庆
  4.陆军军医大学（第三军医大学）军事预防医学系毒理学研究所，重庆
- 作者简介：
  
  LIU Wenbin, E-mail：liuwenbin@tmmu.edu.cn
- 基金信息：
  
  the General National Natural Science Foundation of China(82173556);the Natural Science Foundation of Chongqing(CSTB2024NSCQ-MSX0497);the Key Project of Science and Technology Research Program of Chongqing Education Commission(KJZD-K202512808)
- DOI：10.16016/j.2097-0927.202511095
  CLC： R734.2;R730.23;R363.2
- Received：20 November 2025，
  
  Revised：2026-01-30，
  
  Published：30 March 2026
- 稿件说明：
移动端阅览
邓双武，郭飞，彭万滔，等.苯并（a）芘通过NLRP3焦亡信号通路促进肺癌细胞LTEP-a-2恶性进展：基于生物信息学与细胞实验验证［J］.陆军军医大学学报，2026，48（06）：709-721.DOI：10.16016/j.2097-0927.202511095.

DENG S W，GUO F，PENG W T，et al.Benzo（a）pyrene promotes malignant progression of human lung cancer LTEP-a-2 cells through the NLRP3 pyroptosis signaling pathway： Validation based on bioinformatics and cellular experiments［J］.J Army Med Univ，2026，48（06）：709-721.DOI：10.16016/j.2097-0927.202511095.
邓双武，郭飞，彭万滔，等.苯并（a）芘通过NLRP3焦亡信号通路促进肺癌细胞LTEP-a-2恶性进展：基于生物信息学与细胞实验验证［J］.陆军军医大学学报，2026，48（06）：709-721.DOI：10.16016/j.2097-0927.202511095. DOI：

DENG S W，GUO F，PENG W T，et al.Benzo（a）pyrene promotes malignant progression of human lung cancer LTEP-a-2 cells through the NLRP3 pyroptosis signaling pathway： Validation based on bioinformatics and cellular experiments［J］.J Army Med Univ，2026，48（06）：709-721.DOI：10.16016/j.2097-0927.202511095. DOI：

摘要

目的

苯并（a）芘（benzopyrene，BaP）作为最常见的多环芳烃类污染物，具有强效致癌、致畸和炎症毒性作用，既往研究发现，BaP可通过抑制人支气上皮细胞恶性转化后的铁死亡进而促进其侵袭与迁移能力，但尚不明确BaP能否调控肿瘤细胞焦亡，本研究目的是为了阐明BaP通过抑制LTEP-a-2细胞焦亡促进肺癌侵袭和迁移的作用机制。

方法

基于比较毒理基因组学数据库（comparative toxicogenomics database，CTD）获取BaP可能的潜在基因；基于基因表达数据库（gene expression omnibus，GEO）筛选肺癌相关差异基因。对上述2组基因进行交集分析，获取候选基因集。通过STRING数据库进行蛋白-蛋白相互作用（protein-protein interaction，PPI）网络分析，利用Cytoscape软件进行拓扑属性分析，依据节点度值筛选关键核心靶基因。通过Kaplan-Meier Plotter数据库对PPI网络筛选出的候选基因进行生存分析。采用AutoDock Vina对BaP与各靶蛋白进行分子对接模拟，预测其结合构象并计算结合自由能。使用PyMOL软件进行可视化与分析，通过LigPlot+生成二维相互作用图展示与分析配体-受体间的相互作用。使用梯度浓度BaP（0、0.1、1和10 μmol/L）处理人肺癌细胞LTEP-a-2 48 h，通过CCK-8法检测细胞活力，Transwell实验分析细胞侵袭迁移能力，LDH释放实验评估细胞毒性，采用Annexin V-FITC/PI流式细胞术检测细胞凋亡；通过ELISA定量上清中IL-1β和IL-18水平，采用RT-qPCR和Western blot检测焦亡关键分子（NLRP3、ASC、Caspase-1、GSDMD、IL-1β、IL-18）表达。采用BaP联合焦亡激动剂以及焦亡抑制剂联合焦亡激动剂处理后检测细胞活力与侵袭迁移能力，分析焦亡相关指标（IL-1β/IL-18释放、焦亡关键分子表达）的变化。

结果

体外实验表明，与对照组相比，1和10 μmol/L BaP处理能显著促进人肺癌细胞LTEP-a-2的增殖与侵袭迁移能力（

0.01），但未引起细胞凋亡的显著变化。生物信息学分析提示BaP相关基因显著富集于细胞焦亡通路。不同剂量BaP处理显著抑制了细胞焦亡，表现为LDH释放减少（

0.05），炎症因子IL-1β和IL-18的分泌下调，以及焦亡通路关键分子（NLRP3、ASC、Caspase-1、GSDMD、IL-1β、IL-18）的mRNA和蛋白表达均显著下降（

0.05）。与之相反，焦亡激动剂处理（与对照组相比）则诱发了明显的焦亡表型，并抑制了细胞增殖与侵袭迁移能力（

0.05）。而BaP或焦亡抑制剂联合焦亡激动剂处理组中，焦亡激动剂对肿瘤细胞增殖和侵袭迁移能力的抑制效应均被显著回复（

0.01）。生信预测及分子对接提示NLRP3和PTGS2可能是BaP调控肺癌细胞焦亡的潜在作用靶点。

结论

BaP可通过抑制LTEP-a-2细胞NLRP3-Caspase-1-GSDMD焦亡信号通路，进而促进其侵袭和迁移。

Abstract

Objective

Benzo(a)pyrene (BaP)

as the most common polycyclic aromatic hydrocarbon pollutant

exhibits potent carcinogenic

teratogenic

and inflammatory toxic effects. Previous studies have demonstrated that BaP promotes invasion and migration capabilities of human bronchial epithelial cells following malignant transformation by suppressing ferroptosis; however

whether BaP can regulate tumor cell pyroptosis remains unclear. This study aims to elucidate the mechanism by which BaP promotes lung cancer invasion and migration through suppressing pyroptosis in human lung cancer cell line LTEP-a-2.

Methods

Potential target genes of BaP were retrieved from the Comparative Toxicogenomics Database (CTD)

and lung cancer-related differentially expressed genes (DEGs) were screened from the Gene Expression Omnibus (GEO) database. Subsequently

an intersection analysis of these 2 gene sets was performed to obtain key candidate genes. Protein-Protein Interaction (PPI) network analysis on these candidate genes was conducted using the STRING database

and topological attribute analysis was performed for topological properties with Cytoscape software to screen key hub target genes based on high node degree values. Survival analysis of these PPI-derived hub genes was performed using the Kaplan-Meier Plotter database. Molecular docking simulations between BaP and each target protein were conducted with AutoDock Vina to predict binding conformations and calculate binding free energies. Visualization and analysis of the docking results were carried out using PyMOL software

and detailed ligand-receptor interactions were visualized and analyzed using 2D interaction diagrams generated by LigPlot+. LTEP-a-2 cells were exposed to gradient concentrations of BaP (0

0.1

and 10 μmol/L) for 48 h. Subsequently

cell viability was evaluated with CCK-8 assay

invasive and migratory capabilities were assessed with Transwell assay

cytotoxicity was determined by lactate dehydrogenase (LDH) release assay

and apoptosis was detected

via

Annexin V-FITC/PI flow cytometry. The concentrations of IL-1β and IL-18 in the supernatant were determined by enzyme-linked immunosorbent assay (ELISA). RT-qPCR and Western blotting were performed to measure the expression of key pyroptosis-related molecules (NLRP3

ASC

caspase-1

GSDMD

IL-1β

and IL-18) at mRNA and protein levels. Cell viability and invasive/migratory capabilities were assessed after LTEP-a-2 cells treated with BaP combined with pyroptosis agonists

as well as pyroptosis inhibitors combined with pyroptosis agonists. Concurrently

alterations i

n pyroptosis-related indicators

including IL-1β/IL-18 release and expression of the aforementioned key molecules

were examined.

Results

In vitro

experiments demonstrated that compared to the control group

exposure to 1 or 10 μmol/L BaP significantly promoted the proliferation

invasion

and migration of LTEP-a-2 cells (

0.01)

but did not induced obvious changes in apoptosis. Bioinformatics analysis revealed significant enrichment of BaP-related genes in the pyroptosis pathway. Treatment with different doses of BaP significantly inhibited pyroptosis

as evidenced by reduced LDH release (

0.05)

downregulated secretion levels of the inflammatory cytokines IL-1β and IL-18

and notable decreases in both mRNA and protein expression levels of key molecules in the pyroptosis pathway (NLRP3

ASC

caspase-1

GSDMD

IL-1β

and IL-18) (

0.05). Conversely

pyroptosis agonist (compared to the control group) induced pronounced pyroptotic phenotypes and suppressed cell proliferation and invasive and migratory capacities (

0.05). In the BaP or pyroptosis inhibitor combined with pyroptosis agonist treatment groups

the inhibitory effects of the agonists on tumor cell proliferation and invasion/migration were significantly reversed (

0.01). Bioinformatics prediction and molecular docking analysis indicated that NLRP3 and PTGS2 may serve as potential molecular targets for BaP-mediated regulation of pyroptosis in lung cancer cells.

Conclusion

BaP promotes the invasion and migration of LTEP-a-2 cells by inhibiting the NLRP3-caspase-1-GSDMD pyroptosis signaling pathway.

关键词

Keywords

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