The Effect Of Berberine Combined With Gefitinib On Lung Cancer Based On Network Pharmacology

Objective: The morbidity and mortality rate of lung cancer remains high and become one of the main causes of serious impact on human health.Currently,chemoradiotherapy is the main treatment for lung cancer,and gefitinib is a specific chemotherapy drug in the clinical treatment of lung cancer,acting on EGFR targets,but clinical data show that it has side effects,and patients will develop resistance to it.Berberine extract of Coptis chinensis has great medical value,mainly in the treatment of cardiovascular diseases and digestive diseases,but in recent years,its value against lung cancer has been gradually discovered.Therefore,in this paper,A549 and H1299 cells of non-small cell lung cancer were selected as research objects to explore the effect of berberine combined with gefitinib on non-small cell lung cancer and to study its mechanism of action.Methods: The concentrations of berberine and gefitinib suitable for two kinds of cells were screened by MTT assay.The cells were treated with berberine and gefitinib alone or in combination,and the cell activity was detected by MTT assay.Hoechst staining was used to investigate cell apoptosis.Autophagy was investigated by MDC staining.The cell cycle and apoptosis ratio were detected by flow cytometry.The expression level of related m RNA was detected by q PCR.The common genes of berberine,gefitinib and non-small cell lung cancer were identified by network pharmacology,and PPI protein interaction,GO and KEGG analysis were performed to screen out the possible signaling pathways.Molecular docking was used to detect the binding ability of berberine and receptor protein.Western blot was used to detect protein expression in the signaling pathway.Results: When berberine and gefitinib were combined to treat lung cancer,the cell activity decreased.The number of apoptotic cells increased.Autophagy vesicles and autophagy cells increased.Cell cycle was arrest in G0/G1 phase;The apoptosis rate was increased in A549 and H1299;The m RNA expressions of Bax and LC3-Ⅱwere significantly increased,while the m RNA expressions of Bcl-2 and Cyclin D were significantly decreased.At the same time,network pharmacological exploration found that berberine,gefitinib and non-small cell lung cancer shared five common genes: SRC,KDR,EGFR,MET and EPHB4.Signaling pathways related to the effects of berberine and gefitinib on non-small cell lung cancer mainly include EGFR signaling pathway,MAPK signaling pathway,PI3K-Akt signaling pathway,Ras signaling pathway,etc.Molecular docking showed that berberine and EGFR target could be well combined by HIS-888.After berberine combined with gefitinib,Western blot analysis showed that there was no significant difference in the expression levels of EGFR,PI3 K and AKT,while the expression levels of phosphorylated proteins p-EGFR,p-PI3 K and p-AKT decreased significantly.Conclusion: Berberine combined with gefitinib can inhibit cell proliferation,promote apoptosis and autophagy in non-small cell lung cancer A549 and H1299 cells through EGFR/PI3K/AKT signaling pathway.This study provides theoretical basis and experimental basis for berberine enhancement of gefitinib in the treatment of non-small cell lung cancer,and lays a foundation for the application of berberine in clinical adjuvant treatment of lung cancer.