Abstract
BackgroundNeuroblastoma (NB) is one of most common childhood tumors with high mortality among children worldwide. microRNAs (miRNAs) have been reported to play essential roles in the pathogenesis and therapeutics of NB. However, the role of miR-149 and its mechanism remain poorly understood.Main methodsThe expression levels of miR-149, cell division cycle 42 (CDC42) and B-cell lymphoma 2 (BCL2) were measured in NB tissues or cells by quantitative real-time polymerase chain reaction or western blot. Cell proliferation was measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and colony formation assays. Cell apoptosis was detected by flow cytometry. Chemosensitivity of NB cells to doxorubicin (Dox) was analyzed by MTT assay. The interaction between miR-149 and CDC42 or BCL2 was explored by luciferase activity and RNA immunoprecipitation analyses.ResultsOur data indicated that low expression of miR-149 was displayed in NB tissues and cells and associated with poor survival rate. Overexpression of miR-149 inhibited cell proliferation and colony formation but promoted cell apoptosis and chemosensitivity to Dox in NB cells. Moreover, CDC42 and BCL2 were targeted by miR-149. Additionally, CDC42 and BCL2 mRNA levels were elevated in NB tissues and cells and restoration of CDC42 or BCL2 reversed the regulatory effect of miR-149 on NB progression.ConclusionOur data suggested that miR-149 suppressed cell proliferation and improved Dox chemosensitivity by regulating CDC42 and BCL2 in NB, providing a novel avenue for treatment of NB.
Highlights
Introduction of miR149 facilitates Dox chemosensitivity to NB cells To evaluate the effect of miR-149 on chemosensitivity, SK-N-BE(2)C and SK-N-SH cells transfected with miR-149 or miRNA negative control (miR-NC) were exposed to Dox for 48 h
cell division cycle 42 (CDC42) and B-cell lymphoma 2 (BCL2) mRNA levels were elevated in NB tissues and cells and restoration of CDC42 or BCL2 reversed the regulatory effect of miR-149 on NB progression
Our data suggested that miR-149 suppressed cell proliferation and improved Dox chemosensitivity by regulating CDC42 and BCL2 in NB, providing a novel avenue for treatment of NB
Summary
Introduction of miR149 facilitates Dox chemosensitivity to NB cells To evaluate the effect of miR-149 on chemosensitivity, SK-N-BE(2)C and SK-N-SH cells transfected with miR-149 or miR-NC were exposed to Dox for 48 h. Bioinformatics analysis using starBase showed, among 8 potential targets which were expressed in neuroblastoma, that CDC42 and BCL2 mRNA levels were up-regulated most in SK-N-SH cells (Additional file 1: Figure S1). These two were chosen for further experiments. Up-regulation of miR-149 led to more enrichment of CDC42 and BCL2 by Ago RIP in SK-N-SH and SK-N-BE(2)C cells, whereas little efficacy of enrichment was shown in IgG RIP group (Fig. 4e, f ) These results indicated that CDC42 and BCL2 could be targeted via miR-149 in NB cells. Accruing studies indicate that miR-149 might act as a tumor suppressor in various
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