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Tumour-suppressor microRNAs let-7 and mir-101 target the proto-oncogene MYCN and inhibit cell proliferation in MYCN-amplified neuroblastoma
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Tumour-suppressor microRNAs let-7 and mir-101 target the proto-oncogene MYCN and inhibit cell proliferation in MYCN-amplified neuroblastoma
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Journal Article
Tumour-suppressor microRNAs let-7 and mir-101 target the proto-oncogene MYCN and inhibit cell proliferation in MYCN-amplified neuroblastoma
2011
Overview
Background:
MicroRNAs (miRNAs) regulate expression of many cancer-related genes through posttranscriptional repression of their mRNAs. In this study we investigate the proto-oncogene
MYCN
as a target for miRNA regulation.
Methods:
A luciferase reporter assay was used to investigate software-predicted miRNA target sites in the 3′-untranslated region (3′UTR) of
MYCN
. The miRNAs were overexpressed in cell lines by transfection of miRNA mimics or miRNA-expressing plasmids. Mutation of the target sites was used to validate
MYCN
3′UTR as a direct target of several miRNAs. To measure miRNA-mediated suppression of endogenous N-myc protein, inhibition of proliferation and inhibition of clonogenic growth, miRNAs were overexpressed in a
MYCN
-amplified neuroblastoma cell line.
Results:
The results from this study show that
MYCN
is targeted by several miRNAs. In addition to the previously shown
mir-34a/c
, we experimentally validate
mir-449
,
mir-19a/b
,
mir-29a/b/c, mir-101
and
let-7e/mir-202
as direct
MYCN
-targeting miRNAs. These miRNAs were able to suppress endogenous N-myc protein in a
MYCN
-amplified neuroblastoma cell line. The
let-7e
and
mir-202
were strong negative regulators of
MYCN
expression. The
mir-101
and the let-7 family miRNAs
let-7e
and
mir-202
inhibited proliferation and clonogenic growth when overexpressed in Kelly cells.
Conclusion:
The tumour-suppressor miRNAs
let-7
and
mir-101
target
MYCN
and inhibit proliferation and clonogenic growth of
MYCN
-amplified neuroblastoma cells.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ Biomedical and Life Sciences
/ Down-Regulation - physiology
/ Gene Amplification - physiology
/ Gene Expression Regulation, Neoplastic - genetics
/ Genes
/ Genes, Tumor Suppressor - physiology
/ Humans
/ miRNA
/ mRNA
/ Mutation
/ N-Myc Proto-Oncogene Protein
/ Nuclear Proteins - antagonists & inhibitors
/ Oncogene Proteins - antagonists & inhibitors
/ Oncogene Proteins - genetics
/ Oncology
/ Plasmids
/ Proteins
/ Tumors
