Integrating Transcriptomics and 3D Organoids to Investigate Mechanism of Periplaneta americana Extract Against Lung Adenocarcinoma
10.13422/j.cnki.syfjx.20260261
- VernacularTitle:基于转录组学与3D类器官技术探究美洲大蠊提取物抗肺腺癌的作用机制
- Author:
Qiong MA
1
;
Chunxia HUANG
1
;
Jiawei HE
1
;
Yuting BAI
1
;
Xingyue LIU
1
;
Yuxuan XIONG
1
;
Yang ZHONG
1
;
Hengzhou LAI
1
;
Yuling JIANG
1
;
Xueke LI
1
;
Qian WANG
1
;
Yifeng REN
1
;
Xi FU
1
;
Funeng GENG
2
;
Taoqing WU
2
;
Ping XIAO
3
;
Fengming YOU
1
Author Information
1. Sichuan Provincial Key Laboratory of Traditional Chinese Medicine（TCM） for Prevention and Treatment of Metabolic and Chronic Diseases，Hospital of Chengdu University of TCM，Chengdu 610075，China
2. Sichuan Provincial Key Laboratory of Medicinal Periplaneta americana， Sichuan Good Doctor Panxi Pharmaceutical Co. Ltd.，Chengdu 610031，China
3. Sichuan Cancer Hospital&Institute，Sichuan Clinical Research Center for Cancer， Sichuan Cancer Prevention and Control Center，Afffliated Cancer Hospital of University of Electronic Science and Technology of China，Chengdu 610213，China
- Publication Type:Journal Article
- Keywords:
organoids;
lung adenocarcinoma;
Periplaneta americana;
transcriptome sequencing;
p53 signaling pathway;
breast cancer type 1 susceptibility protein（BRCA1）;
checkpoint kinase 1（CHEK1）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2026;32(11):124-132
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract（PAE） against human-derived lung adenocarcinoma organoids（LUAD-PDOs） and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid（Nor-PDOs） models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8（CCK-8） assay. Experimental groups included the model group（LUAD-PDOs）， normal group， model administration group（LUAD-PDOs+PAE）， and normal administration group（Nor-PDOs+PAE）. Hematoxylin-eosin（HE） staining was used to observe the pathological structures of PDOs， immunohistochemistry（IHC） was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7（CK-7） and Napsin A， TUNEL staining was applied to detect cell apoptosis. RNA sequencing（RNA-Seq） was conducted to identify differentially expressed genes（DEGs）， followed by Gene Ontology（GO）， Kyoto Encyclopedia of Genes and Genomes（KEGG）， and Gene Set Enrichment Analysis（GSEA）， alongside protein-protein interaction（PPI） network analysis to screen core mechanisms. Finally， key targets were validated by integrating external database analysis with immunofluorescence（IF）. ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures， whereas LUAD-PDOs displayed dense， solid structures. CCK-8 and TUNEL assays revealed that， compared with the model group， PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells， while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention（347 up-regulated and 372 down-regulated）（P<0.05）. GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism， including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway（P<0.05）. PPI network analysis indicated that breast cancer type 1 susceptibility protein（BRCA1） and checkpoint kinase 1（CHEK1） were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention（P<0.05）. Furthermore， survival analysis based on The Cancer Genome Atlas（TCGA） database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD（P<0.05）. ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis， its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway， with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.
