The Mechanism of Wenhua Decoction in the Treatment of Non–Small Cell Lung Cancer by Network Pharmacology and Molecular Dynamics Simulation

Introduction

Network pharmacology approaches were applied to reveal the mechanism of Wenhua Decoction in non–small cell lung cancer (NSCLC) treatment.

Methods

Effective chemical constituents and targets of Wenhua Decoction were collected by database, and differentially expressed genes of NSCLC were screened. Restart random walk analysis for protein-protein interaction network was conducted using the intersected genes of differentially expressed genes and target genes in NSCLC as seeds. The top 50 genes in affinity coefficient were used to construct the drug-active constituent-gene interaction network. Molecular docking and molecular dynamic analyses were conducted for core targets and corresponding active constituents in the network. Quercetin content was determined by HPLC. Quantitative real time polymerase chain reaction (qRT-PCR) was performed to assess CDK1 expression in A549 cells. The effects of Wenhua Decoction on the cell cycle were evaluated in sh-CDK1 cells. Cellular thermal shift assay was utilised to examine the binding status between CDK1 and its target quercetin.

Results

The drug-active constituent-gene interaction network demonstrated that CDK1 was at the core of the network. Quercetin could bind to CDK1 stably while causing no significant changes in protein conformation. Wenhua Decoction treatment significantly reduced CDK1 expression and blocked transition of cancer cells from the G1 to the S phase. The quercetin content was 16.0 μg/g. Furthermore, cellular thermal shift assay experiment showed a high binding affinity between CDK1 and quercetin present in the Wenhua Decoction.

Conclusions

This study revealed potential targets, active constituents, and mechanism of Wenhua Decoction in treating NSCLC, providing a reference for clinical treatment of NSCLC with Wenhua Decoction.