Regulation Of MiR-181b On FAMLF In Burkitt Lymphoma/Leukemia

Lymphoma/leukemia is the most common malignant tumor in blood system,mutation and abnormal expression of gene are closely related with the pathoge nesis.Studying the mutation and abnormal expression and regulation mechanism of the related genes can help to elucidate the pathogenesis,to find the specific treatment,to improve the therapeutic effect and survival rate.【Objective】1.To study the correlation between mi R-181 b and FAMLF ge ne expression in Burkitt lymphoma/leukemia.2.To Analyze Whether FAMLF ge ne is the target of mi R-181 b.3.To prove mi R-181 b can directly effect FAMLF gene,down-regulate its expression,inhibit the cell proliferation in Burkitt lym phoma cell line Raji by experiment.【Methods】1.Real-time quantitative PCR was applied to detect the relativ e expression levels of of mi R-181 b and FAMLF in Burkitt lymphoma/leukemia patients,scatter diagram and statistical software SPSS19.0 were applied to anal yze their correlation.2.The bioinformatics software PITA,RNAhybrid,Target Sca n,RNA22 were applied to analyze the target of the mi R-181 b and the effect on FAMLF gene.3.mi R-181 b mimic and negative control substance were transfec ted into the Burkitt lymphomacell line Raji,real-time quantitative PCR was ap plied to detect the express of mi R-181 b,real-time quantitative PCR and Wester n blot were respectively applied to detect the express of FAMLF gene m RNA and protein,MTT assay was applied to detect the proliferation of Raji cell lin e after transfection of mi R-181 b mimic,PI assay was applied to detect the Raj i cell cycle after transfection of mi R-181 b mimic.4.To verify whether mi R-181b has a direct regulation on FAMLF gene,wild type and mutant FAMLF gene Luciferase Report gene vector were respectively constructed,and mi R-181 b an d negative control substance were co-transfected into 293 T cells respectively,th e luciferasegene activity change were observed.【Results】1.Real-time quantitative PCR was applied to detect the relative expression levels of of mi R-181 b and FAMLF in Burkitt lymphoma/leukemia p atients,scatter diagram and SPSS statistical analysis showed that the expression level was significantly negatively related to both.2.Bioinformatics software anal ysis showed that mi R-181 b target 5 ’UTR of FAMLF gene.3.Compared with t he control group,mi R-181 b was significantly increased in the transfected mi R-181 b mimic Raji cell line,FAMLF gene m RNA and protein expression level wa s down-regulated,Raji cell proliferation slowed and cell cycle arrested after tra nsfection,compared with the control group.4.Luciferase report gene experiment al results show that,luciferase report geneactivity was obviously decreased afte r wild type FAMLF gene and mi R-181 b mimic gene were co-transfected into 293 T cells,compared with the control group.【Conclusions】1.Expression of mi R-181 b and FAMLF showed a significa nt negative correlation in Burkitt lymphoma/leukemia patients.2.Mi R-181 b targe ted 5 ’UTR of FAMLF gene.3.Mi R-181 b directly down-regulated FAMLF exp ression and inhibited cell proliferation activity in Burkitt lymphoma Raji cell.