Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025 Apr 25:16:1542116.
doi: 10.3389/fphar.2025.1542116. eCollection 2025.

The mechanistic study of codonopsis pilosula on laryngeal squamous cell carcinoma based on network pharmacology and experimental validation

Huina Guo  1   2 , Yichen Lou  1   2   3 , Xiaofang Hou  1   2   3 , Xiaoya Guan  1   2 , Yujia Guo  1   2 , Qi Han  1   2 , Xuting Xue  1   2 , Ying Wang  1   2 , Long He  1   2 , Zhongxun Li  1   2 , Chunming Zhang  1   2   4
Affiliations

The mechanistic study of codonopsis pilosula on laryngeal squamous cell carcinoma based on network pharmacology and experimental validation

Huina Guo et al. Front Pharmacol. .

Abstract

Introduction: Laryngeal squamous cell carcinoma (LSCC) is a common malignant tumor of the head and neck, with poor prognosis for advanced patients, and there is an urgent need to find new treatment strategies. Codonopsis pilosula, a traditional Chinese medicinal herb, possesses various pharmacological activities, but its antitumor effects and mechanisms in LSCC are still unclear. The aim of this study was to systematically investigate the potential antitumor mechanism of Codonopsis pilosula in LSCC.

Methods: In this study, we screened the effective compounds and targets of Codonopsis pilosula by TCMSP, ETCM and BATMAN-TCM databases, and screened targets related to LSCC by combining DisGeNET, GeneCards database and Cytoscape software. KEGG pathway enrichment analysis was utilized to explore the related signaling pathways. The core targets were further screened based on TCGA and GEO database analysis, and molecular docking was carried out to predict their binding ability to effective compounds. The presence of key compounds was verified by LC-MS, the MAPK3 expression was detected by qPCR in LSCC tissues, and the effects of MAPK3 knockdown on proliferation, migration, invasion, cell cycle, and apoptosis of LSCC cells were evaluated by cellular function assays.

Results: In this study, 22 targets of Codonopsis pilosula that might regulate LSCC were screened based on network pharmacology. KEGG pathway enrichment analysis showed that Codonopsis pilosula-LSCC targets were mainly involved in HIF-1, TNF, IL-17 and FoxO signaling pathways. Based on TCGA and GEO database analysis, MAPK3 was identified as the core target of Codonopsis pilosula-LSCC. The molecular docking results showed that a variety of effective compounds from Codonopsis pilosula had strong binding abilities to MAPK3, among them, Caprylic Acid, Emodin and Luteolin have been confirmed by LC-MS. QPCR analysis indicated that MAPK3 was highly expressed in LSCC tissues. MAPK3 knockdown significantly inhibits LSCC cell proliferation, migration and invasion. It also suppresses LSCC cell growth by blocking the cell cycle and inducing apoptosis.

Conclusion: Codonopsis pilosula exerts antitumor effects in LSCC through the regulation of MAPK3 and multiple signaling pathways, providing a theoretical basis for its clinical application.

Keywords: MAPK3; codonopsis pilosula; experimenntal validation; laryngeal squamous cell carcinoma; network pharmacology.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Article strategy and flowchart.
FIGURE 2
FIGURE 2
Screening of effective compounds and targets for Codonopsis pilosula. (A) Effective compound-target network diagram of Codonopsis pilosula. The green color represents Codonopsis pilosula compounds, the purple and yellow colors represent the corresponding targets of Codonopsis pilosula compounds, and the yellow color represents the 179 key targets of Codonopsis pilosula with a degree >2. (B) Effective compounds with the most targeted interactions in the Codonopsis pilosula effective compounds-targets network diagram.
FIGURE 3
FIGURE 3
Screening of key targets for Codonopsis pilosula in LSCC. (A) Venn diagram showing the intersection of LSCC disease genes obtained from DisGeNET, GeneCards database with Codonopsis pilosula targets. 33 targets were obtained. (B) Based on Cytoscape software topology analysis, targets with degrees>21 were considered key targets. The yellow nodes represent 22 selected key targets.
FIGURE 4
FIGURE 4
MAPK3 as a potential core target for Codonopsis pilosula action in LSCC. (A) The first 30 pathways were mapped according to the proportion of KEGG pathway-enriched genes and p-values. (B) MAPK3 expression was detected by qPCR assay in LSCC and ANM tissues. (C) Classification of compounds identified in Codonopsis pilosula extract by LC-MS analysis. (D) qPCR to detect MAPK3 expression after treatment with FD-LSC-1 and TU-686 at different concentrations of Codonopsis pilosula. Error bars represent the standard deviation (SD) of three independent experiments. *p < 0.05, **p < 0.01.
FIGURE 5
FIGURE 5
Molecular docking and LC-MS total ion chart (TIC) map analysis of effective compounds of Codonopsis pilosula with MAPK3. (A) Molecular docking of effective compounds of Codonopsis pilosula with MAPK3. (B) The compounds TIC of Codonopsis pilosula were identified by LC-MS. Different compounds were detected in positive and negative ion modes, and the chromatographic peaks of the active compounds of interest in the corresponding modes were labeled.
FIGURE 6
FIGURE 6
Knockdown MAPK3 inhibits LSCC cell proliferation, migration and invasion. (A, B) Cell proliferation was determined by CCK8 (A) and RTCA(B) after MAPK3 knockdown in FD-LSC-1 and TU-686 cells. (C–F) Migration (C, D) and invasion (E, F) were examined by transwell assays after MAPK3 silencing in FD-LSC-1 and TU-686 cells. Scale bar, 50 μm. Error bars represent the SD of three independent experiments. *p < 0.05, **p < 0.01.
FIGURE 7
FIGURE 7
Effect of MAPK3 knockdown on apoptosis and cell cycle of LSCC cells. (A–C) The percentage of apoptotic cells was detected by flow cytometry after transfection of FD-LSC-1 and TU-686 cells with si-MAPK3. (D–F) FD-LSC-1 and TU-686 cells were transfected with si-MAPK3, PI staining was performed, and the cell cycle distribution was detected by flow cytometry. Error bars represent the SD of three independent experiments. *p < 0.05, **p < 0.01.
See this image and copyright information in PMC

References

    1. Bhattacharya R., Brown J. S., Gatenby R. A., Ibrahim-Hashim A. (2024). A gene for all seasons: the evolutionary consequences of HIF-1 in carcinogenesis, tumor growth and metastasis. Semin. Cancer Biol. 102–103, 17–24. 10.1016/j.semcancer.2024.06.003 - DOI - PubMed
    1. Cao M., Yu C., Yao Z., Gao X., Wu S. (2019a). Atractylodesin III maintains mitochondrial function and inhibits caspase-3 activity to reverse apoptosis of cardiomyocytes in AMI rats. Int. J. Clin. Exp. Pathol. 12, 198–204. - PMC - PubMed
    1. Cao Y., Dai F., Li Y., Jia L., Luan Y., Zhao Y. (2019b). The research on the mechanism of Tsoong inhibiting for colon cancer. Saudi J. Biol. Sci. 26, 605–613. 10.1016/j.sjbs.2018.11.007 - DOI - PMC - PubMed
    1. Choi S.-H., Kim S.-Y., Kim K.-M., Mony T. J., Bae H. J., Kim M. S., et al. (2023). Fermented sprouts of codonopsis lanceolata suppress LPS-induced inflammatory responses by inhibiting NF-κB signaling pathway in RAW 264.7 macrophages and CD1 mice. Pharmaceutics 15, 1793. 10.3390/pharmaceutics15071793 - DOI - PMC - PubMed
    1. Djoumbou Feunang Y., Eisner R., Knox C., Chepelev L., Hastings J., Owen G., et al. (2016). ClassyFire: automated chemical classification with a comprehensive, computable taxonomy. J. Cheminform 8, 61. 10.1186/s13321-016-0174-y - DOI - PMC - PubMed

LinkOut - more resources