| Objective:This study aims to explore the influence of berberine(BBR)on glioma and to speculate on the mechanism of its effect.Methods:We used CCK-8 to explore the effects on proliferation effects using different concentrations of BBR(6.25,12.5,25,50,100,200μmol/L)on the time of different lengths(24,48,72h)of U251 malignant glioma cells.Subsequently,cell scratch experiment was used to detect the change of scratch area of U251 malignant glioma cells by BBR within 24 hours,and the cell mobility rate was calculated to reflect the effect of migration ability.Then,the TCMSP,Swisstarget Prediction,Batman,Pharm Mapper,and SEA drug target analysis databases were used to find the targets of BBR.The Gene Cards,NCBI,OMIM and CTD disease databases are used to retrieve glioblastoma multiforme(GBM)related targets.We used the online Venn-making software to find the common target genes of BBR and GBM.Next,the common target genes of BBR and GBM were enriched by GO and KEGG,respectively.After processing with R software,the function of the common target genes,their component localization and signaling pathways were found.We used the String to analyze the interactable proteins of BBR and GBM common target genes,and used Cytoscape software to perform visualization and topology analysis.Finally,the relationship between the common target genes and the prognosis of the GBM was analyzed by GEPIA,and the prognosis associated genes were screened.Results:1.CCK-8 assay showed that BBR could effectively inhibit U251 cell activity in the selected concentration range(6.25,12.5,25,50,100,200 μmol/L)and time range(24,48,72 h).The half inhibitory concentration of BBR on U251 cells was 56.08 μmol/L,22.23μmol/L and 14.36 μmol/L within 24,48 and 72 hours,respectively,and there was significant difference in cell activity at different time points(P<0.05).The inhibitory effect of BBR on U251 cell proliferation was positively correlated with time and drug concentration.2.The scratch experiment was observed for 24 hours using 50 μmol / L of BBR,and the results showed that cell mobility of 24 hours > cell mobility of 12 hours > cell mobility of 0 hour both in the control group and the BBR experiment group(P<0.05).At12 and 24 hours,the cell mobility of the control group was higher than the BBR experiment group(P <0.05).3.The target points obtained from the drug database were verified by the Uni Prot database and the duplicates were removed to obtain a total of 201 BBR-related targets.After Gene Cards,NCBI,OMIM and CTD database genes were merged and deleted,3900 GBM-related genes were obtained.After screening by the online Venn-making software,109 common target genes were found.4.The results of enrichment analysis showed that PI3K-Akt signaling pathway,tumor proteoglycan signaling pathway and Rap1 signaling pathway were the main signaling pathways of target genes.The target genes are mainly located on the cell membrane,which are related to cell stress.5.After Cytoscape topology analysis,45 key target genes with scores greater than the average score were selected for the verification of survival analysis.Finally,the differential expression of three genes SOCS3,THBS1 and RAC1 has an impact on the overall survival(log rank P < 0.05).Conclusion:1.BBR can inhibit the proliferation and migration of U251 cells.2.BBR mainly play a role through the PI3K-Akt signaling pathway,and may affect the prognosis of glioma by regulating the expression of SOCS3,THBS1 and RAC1 genes. |
