miRNA -100 promotes proliferation of human leukemia cells HL -60 by targeting carboxy -terminal domain small phosphatase-like protein / 中华实用儿科临床杂志

ABSTRACT

Objective To investigate the effect of miRNA -1 00 on the proliferation of human leukemia cells HL -60,and to explore the mechanism of this action.Methods The bioinformatics software and database were applied to predict and analyze target genes of miRNA -1 00.The vector contained the target gene 3′UTR portion cloned into a luciferase reporter construct.A luciferase reporter assay was performed following co -transfection of small molecular miRNA -1 00 mimics and target gene wild -type or mutant plasmid into HEK -293T cells.HL -60 cells were trans-fected with miRNA -1 00 mimics or anti -miRNA -1 00.After transfection,Western blot was applied to validate the expression of carboxy -terminal domain small phosphatase -like protein (CTDSPL),and the viability of HL -60 was measured by using cell counting kit (CCK -8)assay at 24 h,48 h,72 h,96 h.Results Online software predicted that CTDSPL was likely to be the target gene of miRNA -1 00.Dual luciferase reporter gene assay system showed that miRNA -1 00 could significantly suppress the activity of reporter gene containing CTDSPL 3′-UTR which decreased by about 57.1 %(P =0.000 7).Western blot showed that the expression of CTDSPL was increased after being trans-fected with miRNA -1 00 antisense oligonucleotides and decreased after being transfected with miRNA -1 00 mimics.At the same time,the growth rate of cells treated with miRNA -1 00 mimics or CTDSPL miRNA -1 00 was increased com-pared with that in control by CCK -8 test (P <0.05 ).Conclusions CTDSPL is a downstream target gene of miRNA -1 00.miRNA -1 00 can promote leukemia cell proliferation by inhibiting the expression of CTDSPL.