The Effects Of Mir-490 Family On Biological Behavior Of Bladder Cancer Cell Line EJ And RT4 And Discussing Of Molecular Mechanism

Backgroung:Bladder cancer is one of the most common urologic neoplasms.The prognosis of bladder cancer became poorly when tumor progress to muscular infiltration,the five-year survival rate only about 60-70%.Early diagnosis and treatment is important for delaying progression and survival benefit.The clinical significance is apparent to researching the molecular mechanism of bladder cancer.Recent studies show that microRNAs(miRNAs)play important roles in bladder cancer,some miRNAs may become new tumor marker for diagnosis and disease surveillance.miRNA is one of the non-coding small RNAs contain 21 to 25 bases whicn involve numerous life process and play significant role in post-transcriptional control.This study focus on miR-490 family,designed to research the molecular mechanism and signaling pathway of miR-490 family in bladder cancer.Objectives:Discuss the effects of miR-490 family in tumorigenesis and progression of bladder cancer,research the molecular mechanism and signaling pathway of miR-490 family in bladder cancer.Methods:1.The basal expression level of miR-490 family in EJ and RT4 bladder cancer cell line were screened by quantitative PCR;2.The effects of miR-490 family on cell invasion and cell migration in EJ and RT4 bladder cancer cell line were tested by transwell assay and scratch test;3.The effects of miR-490 family on cell viability in EJ and RT4 bladder cancer cell line were tested by CCK8 assay;4.Bioinformatics methods were used for predicting the possible target genes of miR-490 family;5.After transfection,the expression level change of miR-490 family and possible target genes in EJ and RT4 bladder cancer cell line were screened by quantitative PCR;6.After transfection,the protein level of possible target genes in EJ and RT4 bladder cancer cell line were screened by western blot.Results:1.The basal expression level of miR-490 in RT4 bladder cancer cell line was lower than EJ;2.Transwell assay:miR-490-5p group had higher invasion than control group in both EJ and RT4 cell line,miR-490-3p group had higher invasion and miR-490-3p inhibitor group had lower invasion than control group in RT4 cell line;scratch test:miR-490-5p/3p group had higher migration and miR-490-5p/3p inhibitor group had lower migration than control group in EJ cell line;miR-490-5p/3p inhibitor group had lower migration than control group in RT4 cell line;3.CCK8 assay:miR-490-5p/3p group had higher cell viability than control group in both EJ and RT4 cell line,miR-490-5p/3p inhibitor group had lower cell viability than control group in EJ cell line,miR-490-3p inhibitor group had lower cell viability than control group in RT4 cell line;4.TargetScan 7.1 and miRDB database predicted that the possible target gene of miR-490 family was hnRNPA1;5.Quantitative PCR:miR-490-5p/3p group had higher expression level of miR-490 and miR-490-5p/3p inhibitor group had lower expression level of miR-490 than control group in both EJ and RT4 cell line;miR-490-5p/3p group had higher expression level of hnRNPA1 than control group in EJ cell line,miR-490-5p/3p group had lower and miR-490-5p inhibitor group had higher expression level of hnRNPA1 than control group in RT4 cell line;miR-490-5p/3p group had higher expression level of Kras than control group in both EJ and RT4 cell line;6.Western blot:miR-490 group had higher protein level of Kras and miR-490 had influence on E-cadherin and N-cadherin.Conclusions:1.The basal expression level of miR-490 family is different in various bladder cancer call lines,which may indicate the grade malignancy of tumors;2.miR-490 family promoted the cell viability,cell migration and cell invasion of bladder cancer cells;3.The biological functions of bladder cancer cells were regulated by miR-490 family via hnRNPA1 and Kras;4.miR-490 family had influence on EMT in bladder cancer;5.miR-490 family were oncogenes in bladder cancer.