Effect of fenofibrate on proliferation and apoptosis of human renal transparency cell carcinoma 786-O cells by suppressing STAT3
Objective To explore the role and potential mechanism of fenofibrate in the treatment of renal clear cell carcinoma(ccRCC).Methods The online databases such as Swiss Target Prediction,Pharm Mapper,Target Net,Gene Cards,OMIM,TTD were used to analyze the targets of fenofibrate,ccRCC and apoptosis.The STRING database was applied to construct a protein-protein interaction(PPI)network and visualized using Cytoscape.Molecular function(GO)and pathway(KEGG)enrichment analysis was performed using the Metascapep platform,and Auto Dock Tools and PyMOL software were used for molecular docking.The CCK-8 method was used to detect the effect of fenofibrate on the viability of human renal clear cell carcinoma 786-O cell line and human renal cortical proximal tubular epithelial cell line(HK-2).Scratch assay,clone formation assay,and flow cytometry were used to detect the effects of fenofibrate(50,75,and 100 μmol·L-1)on the migration,proliferation,and cell cycle of 786-O cells,respectively.Hoechst33258 staining and flow cytometry were used to detect the effect of fenofibrate on the apoptosis level of 786-O cells.Western blotting was used to detect the effects of fenofibrate on the expression of signal transduction and transcription activator 3(STAT3),p-STAT3,B-cell lymphoma 2 protein(Bcl-2),Bcl-2 associated X protein(Bax),and Caspase-3 protein in 786-O cells.Results Network pharmacology analysis identified 108 common targets of fenofibrate,ccRCC and apoptosis.Based on the degree values in network topology analysis,the main core targets were selected as STAT3,EGFR,MMP-9,PPARG,RELA and BCL2L.KEGG enrichment analysis revealed that the JAK2-STAT3 signaling pathway played an important role in anti-ccRCC.The molecular docking results showed that fenofibrate had good binding activity with the main core targets STAT3,EGFR,MMP9,PPARG,RELA and BCL2L.The in vitro experimental results showed that compared with the control group,fenofibrate inhibited the growth of 786-O and HK-2 cells in a concentration-and time-dependent manner(P＜0.01,0.001),and had a stronger inhibitory effect on the viability of 786-O cells than HK-2 cells,786-O cell cycle could be arrested in G,phase(P＜0.001),significantly inhibited the colony formation and migration ability of 786-O cells(P＜0.01,0.001),significantly induced apoptosis in 786-O cells(P＜0.001),significantly downregulated p-STAT and Bcl-2,upregulated Bax and Caspase-3 protein expression(P＜0.05).Conclusion Fenofibrate inhibited the proliferation and migration of ccRCC cells,induced apoptosis,and exerted anti-ccRCC effect by downregulation of STAT3.
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