Effects Of Magui Solution On Bladder Cancer T24 Cells And Bioinformatics Analysis Of Its Blood-transmitting Component

Objective: To study the effect of the drug containing serum of Magui liquid on T24 cell line of bladder cancer,analyze its blood components and predict its intervention mechanism.Methods : Firstly,different concentrations of Mag ui liquid containing serum were prepared from SD rats;Using CCK8 experiment to detect the effect of Ma gui liquid containing serum on T24 cell proliferation;Transwell migration test was used to detect the effect of Ma gui liquid containing serum on T24 cell migration;Using Annexin V-APC/PI double staining experiment to detect the effect of Ma gui liquid containing serum on T24 cell apoptosis;Using flow cytometry to detect the effect of Ma gui liquid containing serum on T24 cell cycle;Based on the LC-MS/MS system combined quadrupole Orbitrap mass spectrometer,a comprehensive analysis of the blood components of Ma gui liquid was conducted;The Ch EMBL database and TCMIO database were used to predict the target points of the blood components of Ma gui liquid.GEO database combined with multi chip analysis to obtain the target gene of bladder cancer,Cytoscape＿V3.8.0 software visualization,String database prediction target gene related proteins,use R and related packages to enrich and analyze target genes in GO(Gene Ontology)and KEGG(Kyoto Gene and Genome Encyclopedia).Results: CCK8 experiment results showed that with the increase of drug concentration,the cell viability of bladder cancer T24 cells showed a downward trend.There was no statistical significance between the normal saline group and the blank group(P=0.336),and there was statistical significance between the drug group and the blank group(P<0.001).In the comparison between the drug groups,there was no statistical significance between the normal concentration group,the double concentration group,and the four times concentration group(P values were 0.765 and 0.435,respectively);There was no statistically significant difference between the 2-fold concentration group and the 4-fold concentration group(P=0.628);There was no statistically significant difference between the4-fold concentration group,the 5-fold concentration group,and the7.5-fold concentration group(P values were 0.091 and 0.161,respectively);There was no statistically significant difference in the5-fold concentration group compared to the 7.5 fold concentration group and the 10 fold concentration group(P values were 0.761 and0.056,respectively);The comparison between other drug groups showed statistical significance(P<0.01).Transwell migration experiment results showed that with the increase of drug concentration,the migration number of T24 cells of bladder cancer gradually decreased.All drug groups were statistically significant compared with the blank group(P<0.001),and there was no statistical significance between the 4-fold concentration group and the 5-fold concentration group(P=0.316);There was no statistically significant difference between the 5-fold concentration group and the 7.5-fold concentration group(P=0.062);The comparison between other drug groups showed statistical significance(P<0.01).Annexin V-APC/PI double staining experiment showed that the total apoptosis rate of T24 cells of bladder cancer gradually increased with the increase of Mag ui liquid concentration;The late apoptosis rates of all drug groups were statistically significant compared to the blank group(P<0.001);Except for the normal concentration group and the 4-fold concentration group,the early apoptosis rate of other drug groups was statistically significant compared to the blank group(P<0.01).The results of cell cycle experiment showed that the number of G1 phase cells in the T24 cell cycle of bladder cancer gradually increased with the increase of the concentration of Mag ui liquid,and there was statistical significance between the drug group and the blank group(P<0.001);Except the4-fold concentration group,the number of cells in G2 phase phase in the other drug groups was not statistically significant compared with the blank group;Compared with the blank group,the number of S-phase cells in all drug groups gradually decreased,with statistical significance(P<0.05).The analysis results of the blood components of Ma gui liquid show that there are 71 types of blood components,mainly including phenylpropanoids,flavonoids,phenols,terpenoids,alkaloids,aromatic compounds,and quinones.Bioinformatics analysis results show that there are 18 kinds of active ingredients absorbed by Ma gui liquid,133 corresponding drug targets,770 bladder cancer disease targets,and 13 intersection targets,including TK1,KIF11,CA9,PPARG,BLM,SLC2A1,FEN1,PTGS1,CYP1B1,MAOB,BCL2,ALDH1A1,and MBNL1.The enrichment analysis results of GO and KEGG show that the biological process(BP)with significant enrichment has response to nutrition level,response to oxygen level Response to hypoxia,response to oxygen reduction,and cell response to glucose deficiency.Conclusion: 1.Magui liquid drug containing serum can significantly inhibit the proliferation,migration and cell cycle process of T24 cells of bladder cancer,and promote their apoptosis,and the effect is more obvious with the increase of random drug concentration;2.After entering the bloodstream,the metabolic products of Ma Gui liquid are various,mainly phenylpropanoid,flavonoids,phenols,terpenes,alkaloids,aromatic compounds,and quinones;3.The intervention of Magui liquid on bladder cancer cells is the result of multiple biological processes mediated by multiple targets and the synergistic effect of multiple pathways,mainly involving the transcriptional regulation of hypoxia,energy metabolism,etc;4.Magui liquid has certain anticancer potential and is worthy of further in-depth research.