Screening MiRNAs Related To The Occurrence And Development Of CcRCC Based On Urinary Exosomes,and The Role And Mechanism Of Mir-188-5p In CcRCC

Objective:To select the miRNAs related to the occurrence and development of clear cell renal cell carcinoma(ccRCC)based on urinary exosomes;To validate the role and mechanism of miR-188-5p in ccRCC.Methods and Results:1 The isolation,purification and identification of urinary exosomes.Using ultracentrifugation to isolate and purificate urinary exosomes from ccRCC patients and normal individuals.According to clinical data,the collected samples were divided into vascular invasion group,non-vascular invasion group and normal individual group.2 Screening of miRNAs related to the occurrence of ccRCC.By miRNA sequencing of urinary exosomes,miRNAs significantly differently expressed in non-vascular invasion group vs normal individual group were screened.Subsequently,miRNAs related to the occurrence of ccRCC were selected through single-factor Logistic regression;The model for predicting the occurrence of ccRCC,which had excellent prediction efficiency and could performed independently from clinical data,was established through lasso regression and multi-factor Logistic regression.Through biological enrichment analysis,these miRNAs may be related to ubiquitination-dependent proteolysis and other pathways.3 Screening of miRNAs related to the development of ccRCC.By miRNA sequencing of urinary exosomes,miRNAs significantly differently expressed in vascular invasion group vs non-vascular invasion group were screened.Subsequently,miRNAs related to the prognosis of ccRCC were selected through single-factor COX regression;The model for predicting the prognosis of ccRCC,which had good prediction efficiency and could performed independently from clinical data,was established through multi-factor COX regression and lasso regression.Through biological enrichment analysis,these miRNAs may be related to the connection,activation and transportation of ubiquitin and ubiquitin-like proteins and other pathways.4 Analysis of the role of miR-188-5p in the occurrence and development of ccRCCMiR-188-5p was related to the occurrence and development of ccRCC.Based on TCGA database,it was found that miR-188-5p was associated with higher tumor stage,higher tumor grade and poor prognosis of ccRCC.Subsequently,through single-factor Logistic and COX regression,the target genes of miR-188-5p,which were related to the occurrence and development of ccRCC,were selected.Biological enrichment analysis revealed that these genes may be related to cell movement,immune cells regulation and apoptosis,urogenital system development,and the structure and activity of prominent pathways.At the same time,we also found that PTEN may be the core target gene of miR-188-5p,and there may be two combination sites between them.5 The role and mechanism of miR-188-5p in ccRCCThrough experiments in vitro,we have demonstrated that,comparing to 786-0 cells,the expression of miR-188-5p is higher in CAKI-1 cells,which are more malignant.The ectopic expression of miR-188-5p can inhibit the expression of PTEN,and then increase the phosphorylation of PI3K/AKT-related proteins,thereby promote the proliferation,migration and invasion of renal cancer cells;while the inhibition of miR-188-5p has the opposite effect.Conclusion:1.Based on urinary exosomes,miRNAs differently expressed in non-vascular invasion group vs normal individual group were screened.These miRNAs have good efficiency in predicting the occurrence of ccRCC.They may be related to the ubiquitination-dependent proteolysis.2.Based on urinary exosomes,miRNAs differently expressed in vascular invasion group vs non-vascular invasion group were screened.These miRNAs have good efficiency in predicting the prognosis and development of ccRCC.They may be related to the connection,activation and transport of ubiquitin and ubiquitin-like proteins.3.MiR-188-5p was related to the occurrence and development of ccRCC,and may play a role in regulating immune cells,cell movement and genitourinary system development.4.MiR-188-5p promotes the proliferation,migration and invasion of ccRCC by regulating the PTEN/PI3K/AKT signal axis.