Network pharmacology combined with experimental verification to explore the potential mechanism of naringenin in the treatment of cervical cancer

Abstract

Cervical cancer is the second leading cause of morbidity and mortality in women worldwide. Traditional treatment methods have become limited. Naringenin, a flavonoid abundant in various fruits and herbal medicines, has demonstrated anti-tumor properties among other effects. This research undertook to elucidate the mechanism of naringenin in the context of cervical cancer treatment by leveraging network pharmacology and performing experimental validation. Initial steps involved predicting potential naringenin targets and subsequently screening for overlaps between these targets and those related to cervical cancer, followed by analysis of their interrelationships. Molecular docking was subsequently utilized to verify the binding effect of the central target. Within the framework of network pharmacology, it was discovered that naringenin might possess anti-cancer properties specific to cervical cancer. Following this, the anti-tumor effects of naringenin on Hela cell viability, migration, and invasion were assessed employing CCK-8, transwell, wound healing assays, and western blotting. Experimental data indicated that naringenin attenuates the migration and invasion of Hela cells via downregulation EGFR/PI3K/AKT signaling pathway. Thus, our findings suggest that naringenin has therapeutic impacts on cervical cancer via multiple mechanisms, primarily by inhibiting the migration and invasion through the EGFR/PI3K/AKT/mTOR pathway. This study offers fresh insights for future clinical studies.

Figure 1

Flowchart of the research process.

Figure 2

Predictive targets of naringenin and cervical cancer. (A) Identification of differentially expressed genes (DEGs) in cervical cancer versus normal cervical tissues. Downregulated genes are represented in blue and upregulated genes in red. (B) The Venn diagram illustrating common targets in cervical cancer and naringenin. (C) Targets of naringenin relevant to cervical cancer.

Figure 3

Protein–protein interaction (PPI) network. (A) PPI network of naringenin-cervical cancer targets. (B) Hub targets of naringenin relevant to cervical cancer.

Figure 4

Enrichment analysis of GO and KEGG pathways. (A–C) Top 20 of GO enrichment analysis for potential targets. (D) Top 20 “target-pathways against cervical cancer” network.

Figure 5

Compound targets subjected to detailed docking simulations with four highest molecular docking affinities (A) MMP9, (B) EGFR, (C) ESR1, (D) AKT1.

Figure 6

Impact of naringenin on morphology and viability of HeLa cells. (A) Microscopic imaging of cell morphology at 200× magnification after incubation with varied concentrations of naringenin for 24 h. (B) Viability rates assessed by the CCK-8 assay. Data from three independent experiments are presented as means and standard deviations. *p < 0.05.

Figure 7

Impact of naringenin on migration and invasion of cervical cancer cells. (A) Cell migration assessed through wound healing assay. (B) Cell invasion evaluated by Transwell assay. Data are expressed as mean ± SD. *p < 0.05; **p < 0.01.

Figure 8

Inhibition of naringenin on EGFR/PI3K/AKT/mTOR pathway. Cropped blots were shown in Fig. 8, and the original blots were presented in Supplementary Fig. 1. Protein expression levels of EGFR, PI3K, p-PI3K, AKT, p-AKT, mTOR, and caspase-3, as detected via western blotting.

Figure 9

Schematic diagram of naringenin in anti-cervical cancer.