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Journal of Experimental & Clinical Cancer Research

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Open Access 01-12-2011 | Research

miR-15a and miR-16-1 inhibit the proliferation of leukemic cells by down-regulating WT1 protein level

Authors: Shen-meng Gao, Chong-yun Xing, Chi-qi Chen, Si-si Lin, Pei-hong Dong, Fu-jun Yu

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2011

Abstract

Background

miR-15a and miR-16-1(miR-15a/16-1) have been implicated as tumor suppressors in chronic lymphocytic leukemia, multiple myeloma, and acute myeloid leukemic cells. However the mechanism of inhibiting the proliferation of leukemic cells is poorly understood.

Methods

K562 and HL-60 cells were transfected with pRS-15/16 or pRS-E, cell growth were measured by CCK-8 assay and direct cell count. Meanwhile WT1 protein and mRNA level were measured by Western blotting and quantitative real-time PCR.

Results

In this study we found that over-expression of miR-15a/16-1 significantly inhibited K562 and HL-60 cells proliferation. Enforced expression of miR-15a/16-1 in K562 and HL-60 cells significantly reduced the protein level of WT1 but not affected the mRNA level. However enforced expression of miR-15a/16-1 can not reduce the activity of a luciferase reporter carrying the 3'-untranslated region(3'UTR) of WT1. Silencing of WT1 by specific siRNA suppressed leukemic cells proliferation resembling that of miR-15a/16-1 over-expression. Anti-miR-15a/16-1 oligonucleotides (AMO) reversed the expression of WT1 in K562 and HL-60 cells. Finally, we found a significant inverse correlation between miR-15a or miR-16-1 expression and WT1 protein levels in primary acute myeloid leukemia (AML) blasts and normal controls.

Conclusions

These data suggest that miR-15a/16-1 may function as a tumor suppressor to regulate leukemic cell proliferation potentially by down-regulating the WT1 oncogene. However WT1 is not directly targeted by miR-15a/16-1 through miRNA-mRNA base pairing, therefore more study are required to understand the mechanism by which miR-15a/16-1 downregulate WT1.

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Title: miR-15a and miR-16-1 inhibit the proliferation of leukemic cells by down-regulating WT1 protein level
Authors: Shen-meng Gao
Chong-yun Xing
Chi-qi Chen
Si-si Lin
Pei-hong Dong
Fu-jun Yu
Publication date: 01-12-2011
Publisher: BioMed Central
Published in: Journal of Experimental & Clinical Cancer Research / Issue 1/2011
Electronic ISSN: 1756-9966
DOI: https://doi.org/10.1186/1756-9966-30-110

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Background

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Conclusions

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