MiRNA-100 promotes proliferation of human leukemia cells HL-60 by targeting carboxy-terminal domain small phosphatase-like protein

Abstract

Objective To investigate the effect of miRNA-100 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-100. 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-100 mimics and target gene wild- type or mutant plasmid into HEK-293T cells. HL-60 cells were transfected with miRNA-100 mimics or anti-miRNA-100. 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-100.Dual luciferase reporter gene assay system showed that miRNA-100 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 transfected with miRNA-100 antisense oligonucleotides and decreased after being transfected with miRNA-100 mimics.At the same time, the growth rate of cells treated with miRNA-100 mimics or CTDSPL miRNA-100 was increased compared with that in control by CCK-8 test(P<0.05). Conclusions CTDSPL is a downstream target gene of miRNA-100.miRNA-100 can promote leukemia cell proliferation by inhibiting the expression of CTDSPL. Key words: miRNA-100; Carboxy-terminal domain small phosphatase-like protein; Proliferation