关键词: 脑胶质瘤, 洛莫司汀, 分子动力学模拟, 分子对接

Abstract: Objective Based on methods such as network pharmacology,molecular docking and molecular dynamic simulation,this study aimed to explore the potential targets and molecular mechanism of lomustine in the treatment of glioma,providing a theoretical basis for clinical precision medicine. Methods Potential drug targets were obtained by searching SwissTargetPrediction,Comparative Toxicogenomics database,and Genecards databases.Protein-protein interaction(PPI)networks were constructed through the STRING database to screen core targets,and perform GO functional enrichment analysis on the core targets.Molecular docking technology was used to predict the binding mode and binding energy between lomustine and core targets,and evaluate the stability of target binding through molecular dynamics simulation and related methods.Differential expression and survival analysis were performed using the GEPIA database,and the correlation between the final core target genes and overall survival(OS)and disease-free survival(DFS)of patients was assessed.Finally,in vitro experiments were performed to evaluate the effects of targetCDKN1A knockdown and lomustine treatment on the biological behaviors of the human glioma U-87MG cells.RT-qPCR and Western blot were used to detect the levels of CDKN1A knockdown at mRNA and protein expression in U-87MG cells.Wound healing assay was conducted to evaluate cell migration ability;CCK-8 assay was used to assess cell proliferation ability;Flow cytometry was performed to detect cell apoptosis in U-87MG cells. Results As a result of screening,113 potential targets of lomustine and 3 935 targets associated with glioma were identified,and 10 candidate targets were obtained by taking the intersection.Six core targets were identified through PPI network screening,followed by further screening of five final analytical targets through molecular docking.Molecular dynamic simulation ultimately determined CDKN1A as the core target of action.This target was highly expressed in glioma tissues and was associated with poor OS and DFS(P＜0.05),mainly localized in the nucleus.knockdown of CDKN1A gene and lomustine treatment could significantly inhibit cell migration and proliferation(P＜0.01),and promoted apoptosis(P＜0.05)in U-87MG cells,and their effects were comparable. Conclusion CDKN1A may be as a key potential target for the treatment of glioblastoma with lomustine,providing new theoretical basis for further elucidating the anti-glioma mechanism and clinical application of lomustine.

Key words: glioma, lomustine, molecular dynamics simulation, molecular docking