Inhibitory effect of Fuzheng Huaji Decoction against non-small cell lung cancer cells in vitro and the possible molecular mechanism

doi:10.12122/j.issn.1673-4254.2025.06.04

Abstract

Abstract:

Objective To investigate the inhibitory effect of Fuzheng Huaji Decoction against non-small cell lung cancer (NSCLC) cells in vitro and explore the underlying mechanism. Methods The active ingredients and targets of Fuzheng Huaji Decoction were identified using TCMSP and SwissTargetPrediction databases. NSCLC-related targets from GeneCards and PharmGKB were intersected with the targets of the Decoction, and a protein-protein interaction (PPI) network was constructed to identify the core targets, which were analyzed with GO and KEGG pathway enrichment analysis. Cultured A549 cells were treated with different concentrations of Fuzheng Huaji Decoction-medicated serum, and the changes in cell proliferation, apoptosis, and protein expressions were examined using CCK-8 assay, annexin V-FITC/PI staining and Western blotting. Results Fuzheng Huaji Decoction contained 140 active ingredients, and 707 drug-disease intersecting targets were identified. Among these targets, TP53, AKT1, HIF1A, GAPDH, ALB, EGFR, CTNNB1, and TNF were identified as the core targets which were involved in the biological processes related to kinases and receptors and the PI3K-AKT, Ras, calcium, and MAPK pathways. Molecular docking studies indicated strong binding affinity of the active ingredients with TP53, AKT1, and HIF1A. In cultured A549 cells, treatment with 2.5%, 5%, and 10% Fuzheng Huaji Decoction-medicated serum significantly inhibited cell proliferation, promoted cell apoptosis, and downregulated the expression levels of HIF1A, p-AKT (Thr308), and TP53 proteins. Conclusion Fuzheng Huaji Decoction inhibits proliferation of NSCLC cells possibly by downregulating the expressions of HIF1A, p-AKT (Thr308), and TP53.

Key words: Fuzheng Huaji Decoction, non-small cell lung cancer, network pharmacology, molecular docking, in vitro experiments

Lijun HE, Xiaofei CHEN, Chenxin YAN, Lin SHI. Inhibitory effect of Fuzheng Huaji Decoction against non-small cell lung cancer cells in vitro and the possible molecular mechanism[J]. Journal of Southern Medical University, 2025, 45(6): 1143-1152.

Figures/Tables 11

Fig.1 Venn diagram of drug targets of Fuzheng Huaji Decoction and NSCLC disease targets.

Fig.2 Component-target-disease map.

Tab.1 Key active components of Fuzheng Huaji Decoction for treatment of non-small cell lung cancer

Code	ID	Name	Relevance	Source
DZ8	MOL006709	8-Hydroxypinoresinol	149	Eucommia
FBX3	MOL002714	baicalein	83	Pinellia
HQ2	MOL000239	Jaranol	81	Astragalus
DS10	MOL000006	luteolin	80	Codonopsis
DZ25	MOL000098	quercetin	80	Eucommia
DZ4	MOL000422	kaempferol	80	Eucommia
HQ15	MOL000422	kaempferol	80	Astragalus
HQ20	MOL000098	quercetin	80	Astragalus
HQ5	MOL000354	isorhamnetin	80	Astragalus
HQ8	MOL000378	7-O-methylisomucronulatol	80	Astragalus

Fig.3 Target protein-protein interaction network analysis of Fuzheng Huaji Decoction for treatment of NSCLC.

Fig.4 GO function enrichment analysis.

Fig.5 KEGG pathway enrichment analysis.

Tab.2 Binding energies of the core active components of Fuzheng Huaji Decoction for NSCLC treatment for docking with the core target molecules TP53, AKT1, and HIF1A

Target	Sample	Estimated ΔG (kcal/mol)
TP53	DZ8	-9.40
TP53	FBX3	-9.90
TP53	HQ2	-9.50
TP53	DS10	-10.40
TP53	DZ25	-10.20
TP53	DZ4	-9.80
TP53	HQ5	-9.60
TP53	HQ8	-8.70
AKT1	DZ8	-10.80
AKT1	FBX3	-10.20
AKT1	HQ2	-9.70
AKT1	DS10	-11.10
AKT1	DZ25	-9.70
AKT1	DZ4	-9.70
AKT1	HQ5	-9.50
AKT1	HQ8	-9.40
HIF1A	HQ8	-4.80
HIF1A	DZ8	-5.50
HIF1A	HQ2	-4.90
HIF1A	HQ5	-5.00
HIF1A	FBX3	-5.40
HIF1A	DZ4	-4.90
HIF1A	DZ25	-5.10
HIF1A	DS10	-6.00

Fig.6 Visualization of the results of molecular docking analysis. A: Docked molecular conformation of TP53 with DS10; B: Molecular conformation of AKT1 docked with DS10; C: Molecular conformation of HIF1A docked with DS10.

Fig.7 Changes in proliferation of A549 cells treated with Fuzheng Huaji Decoction-medicated serum detected by CCK-8 assay. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs control group.

Fig.8 Apoptosis of A549 cells treated with Fuzheng Huaji Decoction-medicated serum detected by annexin V-FITC/PI staining. **P<0.01, ***P<0.001 vs TCRL.

Fig.9 Effect of Fuzheng Huaji Decoction-medicated serum on expressions of HIF1A, p-AKT 308 and TP53 proteins in A549 cells detected by Western blotting proteins in A549 cells. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 vs NCgroup.

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