Experimental Filtering Of MicroRNAs Targeting On The 3’UTR Of Human TCP1 Gene

T-complex 1(TCP1),α subunit of chaperonin CCT,was significantly relative to the increased malignancy,enhancing proliferation and invasiveness of HL60 cell line.Our previous results showed that TCP1 plays an important role in the development of acute leukemia.In recent years,people put eyes on the acting of miRNA on tumorigenesis increasingly.Researches focusing on miRNA and tumor have been valued,and scholars come to committing to find new diagnostic criteria and therapeutic targets for tumors.Experimental filtering of micro RNAs targeting on the 3’Untranslated Region(3’UTR)of human TCP1 gene may have significance for diagnosis and treatment of acute leukemia.Objective: Experimental filtering of micro RNAs targeting on the 3’UTR of human TCP1 gene.Methods: Molecular cloning techniques were used to construct the dual luciferase reporter plasmid which contained the 3’UTR of human TCP1 gene.Six tested miRNAs(hsa-miR-142-5p,hsa-mir-33b-5p,hsa-miR-340-5p,hsa-miR-3143,hsa-miR-543 and hsa-miR-429),which were confirmed that relative to PI3K/AKT signaling pathway or generation and progression of some tumors,were selected by bioinformatics software combining with published papers.The recombinant plasmid and mimics were transfected into 293 T cells using liposomes.Fluorescence and protein were tested to detect which miRNA could target on TCP1 3’UTR and regulate the expression of TCP1.Results: The sequencing results showed the success of recombining the plasmid and mutant plasmid.After twenty-four hours,cotransfecting mimic of hsa-miR-340-5p and recombined plasmid group showed lower fluorescence;transfecting mimic of hsa-miR-340-5p group showed reduced TCP1 repression.Conclusion: MiRNA hsa-miR-340-5p could target regulating the expression of TCP1.For TCP1 is significantly related to acute leukemia,miRNA hsa-miR-340-5p may related to acute leukemia as well.There are more depth study should be done to find meaningful diagnostic indicators and therapeutic targets.