Integrating Transcriptomics and 3D Organoids to Investigate Mechanism of <italic style="font-style: italic">Periplaneta americana</italic> Extract Against Lung Adenocarcinoma

MA Qiong; HUANG Chunxia; HE Jiawei; BAI Yuting; LIU Xingyue; XIONG Yuxuan; ZHONG Yang; LAI Hengzhou; JIANG Yuling; LI Xueke; WANG Qian; REN Yifeng; FU Xi; GENG Funeng; WU Taoqing; XIAO Ping; YOU Fengming

doi:10.13422/j.cnki.syfjx.20260261

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Integrating Transcriptomics and 3D Organoids to Investigate Mechanism of Periplaneta americana Extract Against Lung Adenocarcinoma

更新时间：2026-04-30

- Integrating Transcriptomics and 3D Organoids to Investigate Mechanism of Periplaneta americana Extract Against Lung Adenocarcinoma
  增强出版
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 32, Issue 11, Pages: 124-132(2026)
- 作者机构：
  
  1.成都中医药大学附属医院，代谢与慢病中医药防治四川省重点实验室，成都 610075
  2.成都中医药大学肿瘤研究所，成都 610032
  3.四川好医生攀西药业有限责任公司，药用美洲大蠊四川省重点实验室，成都 610031
  4.电子科技大学附属肿瘤医院，四川省肿瘤医院（研究所），四川省肿瘤临床医学研究中心，四川省癌症防治中心，成都 610213
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20260261
  CLC： R242;R730.5;R273
- Received：26 November 2025，
  
  Revised：2026-01-16，
  
  Accepted：25 February 2026，
  
  Online First：02 March 2026，
  
  Published：05 June 2026
- 稿件说明：
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马琼,黄春霞,何佳玮等.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[J].中国实验方剂学杂志,2026,32(11):124-132.

MA Qiong,HUANG Chunxia,HE Jiawei,et al.Integrating Transcriptomics and 3D Organoids to Investigate Mechanism of Periplaneta americana Extract Against Lung Adenocarcinoma[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(11):124-132.
马琼,黄春霞,何佳玮等.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[J].中国实验方剂学杂志,2026,32(11):124-132. DOI： 10.13422/j.cnki.syfjx.20260261.

MA Qiong,HUANG Chunxia,HE Jiawei,et al.Integrating Transcriptomics and 3D Organoids to Investigate Mechanism of Periplaneta americana Extract Against Lung Adenocarcinoma[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(11):124-132. DOI： 10.13422/j.cnki.syfjx.20260261.

摘要

目的

评估美洲大蠊提取物（PAE）对人源性肺腺癌类器官（LUAD-PDOs）的抗肿瘤作用，并基于转录组学阐明其作用机制。

方法

收集LUAD患者的肿瘤及癌旁正常新鲜组织，采用3D类器官培养技术构建LUAD-PDOs及正常肺类器官（Nor-PDOs）。实验分为正常组、正常给药组（Nor-PDOs+PAE）、模型组及模型给药组（LUAD-PDOs+PAE）。通过细胞增殖与活性检测（CCK-8）法确定PAE的有效干预浓度。采用苏木素-伊红（HE）染色观察类器官病理学结构；免疫组化（IHC）检测核增殖抗原Ki-67及肺腺癌分化标志物细胞角蛋白-7（CK-7）、天冬氨酸蛋白酶A（Napsin A）的表达；原位末端标记法（TUNEL）检测细胞凋亡程度；转录组测序（RNA-Seq）筛选差异表达基因（DEGs），并进行基因本体（GO）功能分析、京都基因和基因组百科全书（KEGG）通路富集分析、基因集富集分析（GSEA）及蛋白质-蛋白质相互作用（PPI）网络筛选核心机制。最后，联合外部数据及免疫荧光（IF）对关键靶点进行验证。

结果

成功建立了病理特征与原发组织高度一致的LUAD-PDOs及Nor-PDOs。CCK-8确定PAE的有效干预质量浓度为16 g·L

-1

。形态学观察显示，Nor-PDOs表现出空腔结构，而LUAD-PDOs呈致密、实性结构。CCK-8及TUNEL检测发现，与模型组比较，PAE干预抑制了LUAD-PDOs的增殖，并促进LUAD细胞凋亡，而PAE对Nor-PDOs的活性差异无统计学意义。转录组分析鉴定出PAE干预后逆转了719个DEGs表达（上调347个，下调372个）（

0.05）。GO富集分析显示，与模型组比较，模型给药组的DEGs明显富集于细胞周期调控相关生物学过程；KEGG通路分析揭示，PAE可调控肿瘤发生、p53信号通路等增殖代谢相关通路；GSEA进一步证实PAE可明显促进p53信号通路的活性（

0.05）；PPI网络分析表明乳腺癌易感蛋白1（BRCA1）与检查点激酶1（CHEK1）为p53通路中的核心下调靶点；IF验证了BRCA1与CHEK1在LUAD-PDOs中高表达，并在PAE干预后表达明显降低。此外，癌症基因组图谱（TCGA）数据库生存分析表明，BRCA1与CHEK1低表达与LUAD患者的总生存期延长明显相关（

0.05）。

结论

PAE可有效抑制LUAD-PDOs的增殖并促进其凋亡，其抗肿瘤机制与p53信号通路活性相关，BRCA1与CHEK1基因可能是PAE发挥作用的关键下游靶点。

Abstract

Objective

To 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.

Methods

Fresh 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）.

Results

Nor-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 d

own-regulated）（

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（

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（

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（

0.05）.

Conclusion

PAE 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.

关键词

Keywords

references

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Integrating Transcriptomics and 3D Organoids to Investigate Mechanism of Periplaneta americana Extract Against Lung Adenocarcinoma

DOI：10.13422/j.cnki.syfjx.20260261

摘要

Abstract

关键词

Keywords

references