Exploration Of Related Genes In Renal Tubular Injury Based On Bioinformatics

Objective: to find out the genes that may play a role in renal tubular injury by mining the kidney disease related database and analyzing the existing data based on bioinformatics.The relationship between these genes and renal tubular injury was discussed by some basic experiments.Methods:(1)data mining related to nephropathy was carried out by GEO(https://www.ncbi.nlm.nih.gov/geo/),String(https://string-db.org/)protein interaction website,Coexpedia(http://www.coexpedia.org/)gene interaction website,GO function enrichment analysis of Cytosacpe,KEGG(https://www.kegg.jp/)pathway analysis and nephropathy clinical database(https://www.nephroseq.org))? Four key genes with high scores were obtained,including CDC25 C and MAD2L1 involved in cell cycle(hsa04110),BIRC5 involved in apoptosis(hsa04210),EXO1 involved in DNA mismatch repair(hsa03430).Finally,CDC25 C and MAD2L1 were selected as the research objects for further experiments.(2)Cisplatin was used to intervene HK-2 in renal tubular cells.The concentrations of cisplatin were 3.75ug/ml,7.5ug/ml,15 ug / ml.The model of renal tubule injury was established by detecting the changes of cell morphology,cell nucleus and CCK8 cell growth inhibition.Then,RNA was extracted from different degrees of renal tubular injury model,RT-PCR was carried out to detect the expression of CDC25 C and MAD2L1;CDC25C and MAD2L1 inhibitors were used to intervene their expression,CCK8 was used to detect the growth inhibition of cells,and the effect of CDC25 C and MAD2L1 expression on the degree of renal tubular injury was evaluated.Results:(1)compared with the normal control group,cisplatin intervention model cell morphology and nuclear morphology changed significantly,cell survival rate decreased,renal tubular injury model was successfully established.The cell survival rates of 3.75ug/ml,7.5ug/ml and 15ug/ ml were 83.5%,56.1% and 21.1%,respectively.The half inhibitory concentration of cisplatin was about 7.5ug/ml.Therefore,cisplatin concentration of 7.5 was used as moderate injury group,cisplatin concentration of 3.75 and cisplatin concentration of 15 were used as mild injury group and severe injury group for further study.(2)The expression of CDC25 C and MAD2L1 was detected by RT-PCR.It was found that the expression of CDC25 C and MAD2L1 increased gradually with the severity of renal tubular injury.After inhibiting the expression of CDC25 C and MAD2L1 with inhibitors in cisplatin injury model.CCK8 was used to detect the cell survival.It was found that the cell survival rate of MAD2L1 inhibition group decreased significantly in mild,medium and severe injury model.There was no significant difference in the cell survival rate of CDC25 C inhibition group in mild and moderate injury,but in severe injury group,the cell survival rate of the inhibition group increased.Conclusion: In the renal tubular injury,the expression of CDC25 C and MAD2L1 increased gradually when the degree of injury increased.Inhibition of the expression of MAD2L1 in renal tubular injury leads to more severe tubular injury.