Effects Of Monomeric Euphorin A From Jingjingji On The Proliferation And Apoptosis Of Human Breast Cancer Cells MDA-MB-231

Objective: To analyze the potential mechanism of Euphorbia pekinensis in the treatment of breast cancer by using the method of network pharmacology,and observe the effect of proliferation,cell cycle and apoptosis of breast cancer cell MDA-MB-231 by the monomer compound Ebracteolatain A(EA)of Euphorbia pekinensis by cell experiments.Material and method:1.The compound components and related targets of Euphorbia pekinensis were retrieved by using the traditional Chinese medicine system pharmacology database and analysis platform(TCMSP).The Gend Cards and OMIM databases were used to screen breast cancer disease targets,and a visual network diagram of "drugs-key compounds-disease targets" was constructed with Cytoscape(3.7.2)software.The PPI protein interaction network was constructed by String database platform,and then the Bioconductor platform was used for GO enrichment analysis and pathway enrichment analysis.2.The CCK-8 experiment method was used to detect whether Ebracteolatanolide A has an effect on the proliferation of human breast cancer MDA-MB-231 cells.The concentration of Ebracteolatanolide A was 5mmol/L,10mmol/L,20mmol/L,40mmol/L,60mmol/L,80mmol/L,100mmol/L and calculate the IC50 value;Flow cytometry was used to detect the effect of Ebracteolatanolide A(50mmol/L,100mmol/L)on the cell cycle and apoptosis of human breast cancer MDA-MB-231 cells.Results:1.A total of 7 key compounds related to disease targets were obtained,which mainly play a role in the treatment of breast cancer by regulating target genes such as GSK3B、ESR1、MAPK14、PTGS2、ADRB2、AR、ESR2,and estrogen signaling pathway、thyroid hormone signaling pathway、prolactin signaling pathway、breast cancer pathway and so on.2.Ebracteolatanolide A can inhibit the proliferation of human breast cancer MDA-MB-231 cells and MCF-7 cells at different concentrations and at different times,showing a clear time and concentration dependence.The inhibitory effect of the EA on MDA-MB-231 cells was more obvious than that of MCF-7 cells: the IC5048 h of MDA-MB-231 was 566.88μmol/L,IC5072 h was 106.76μmol/L;the IC5048 h of MCF-7 was 844.32μmol/ L,IC5072 h was362.43μmol/L.(P<0.05).3.MDA-MB-231 cells were treated with 50μmol/L and 100μmol/L Ebracteolatanolide A for72 h and then tested by flow cytometry.The results showed that the proportion of cells in G0/G1 phase decreased,and the proportion of cells in G2/M phase increased(P< 0.05),the cells are arrested in G2/M phase.4.MDA-MB-231 cells were treated with 50μmol/L and 100μmol/L Ebracteolatanolide A for72 h,and the results were detected by flow cytometry.The results showed that Ebracteolatanolide A could induce the apoptosis of MDA-MB-231 cells,and the apoptosis of MDA-MB-231 cells was mainly induced at terminal stage.Conclusion:1.Based on the analysis of network pharmacology,the results suggested Euphorbia pekinensis play a vital role in inhibiting breast cancer with multiple components,multiple targets,and multiple pathways.2.Ebracteolatanolide A had a significant inhibitory effect on cell proliferation of MDA-MB-231 cells and MCF-7 cells at final concentrations from 50μmol/L to 100μmol/L for 48 h and 72 h,and the inhibitory effect was also time-dependent inhibition.And the inhibitory effect of the EA on MDA-MB-231 cells was more significant than that of MCF-7 cells.3.After treatment of Ebracteolatanolide A with different concentrations,cell cycle arrest occurred in MDA-MB-231 breast cancer cells,and the arrest mainly appeared in G2/M phase.4.Ebracteolatanolide A can induce cell apoptosis with diverse concentrations,and mainly induce cell apoptosis in the late stage.