In hepatocellular carcinoma miR-519d is up-regulated by p53 and DNA hypomethylation and targets CDKN1A/p21, PTEN, AKT3 and TIMP2

Francesca Fornari; Maddalena Milazzo; Pasquale Chieco; Massimo Negrini; Elena Marasco; Giovanni Capranico; Vilma Mantovani; Jessica Marinello; Silvia Sabbioni; Elisa Callegari; Matteo Cescon; Matteo Ravaioli; Carlo M Croce; Luigi Bolondi; Laura Gramantieri

doi:10.1002/path.3995

In hepatocellular carcinoma miR-519d is up-regulated by p53 and DNA hypomethylation and targets CDKN1A/p21, PTEN, AKT3 and TIMP2

J Pathol. 2012 Jul;227(3):275-85. doi: 10.1002/path.3995. Epub 2012 Apr 18.

Authors

Francesca Fornari¹, Maddalena Milazzo, Pasquale Chieco, Massimo Negrini, Elena Marasco, Giovanni Capranico, Vilma Mantovani, Jessica Marinello, Silvia Sabbioni, Elisa Callegari, Matteo Cescon, Matteo Ravaioli, Carlo M Croce, Luigi Bolondi, Laura Gramantieri

Affiliation

¹ Centro di Ricerca Biomedica Applicata, Policlinico S. Orsola-Malpighi e Università di Bologna, Italy.

PMID: 22262409
DOI: 10.1002/path.3995

Abstract

MiR-519d belongs to the chromosome 19 miRNA cluster (C19MC), the largest human miRNA cluster. One of its members, miR-519d, is over-expressed in hepatocellular carcinoma (HCC) and we characterized its contribution to hepatocarcinogenesis. In HCC cells, the over-expression of miR-519d promotes cell proliferation, invasion and impairs apoptosis following anticancer treatments. These functions are, at least in part, exerted through the direct targeting of CDKN1A/p21, PTEN, AKT3 and TIMP2. The mechanisms underlying miR-519d aberrant expression in HCC were assayed by genomic DNA amplification, methylation analysis and ChIP assay. The aberrant hypomethylation of C19MC and TP53 were respectively identified as an epigenetic change allowing the aberrant expression of miR-519d and one of the factors able to activate its transcription. In conclusion, we assessed the oncogenic role of miR-519d in HCC by characterizing its biological functions, including the modulation of response to anticancer treatments and by identifying CDKN1A/p21, PTEN, AKT3 and TIMP2 among its targets.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Aged
Aged, 80 and over
Antibiotics, Antineoplastic / pharmacology
Apoptosis
Carcinoma, Hepatocellular / enzymology*
Carcinoma, Hepatocellular / genetics
Carcinoma, Hepatocellular / pathology
Cell Cycle / drug effects
Cell Movement / drug effects
Cell Proliferation / drug effects
Chromatin Immunoprecipitation
Cyclin-Dependent Kinase Inhibitor p21 / genetics
Cyclin-Dependent Kinase Inhibitor p21 / metabolism*
DNA Methylation*
Doxorubicin / pharmacology
Female
Gene Expression Regulation, Neoplastic / drug effects
Hep G2 Cells
Humans
Liver Neoplasms / enzymology*
Liver Neoplasms / genetics
Liver Neoplasms / pathology
Male
MicroRNAs / metabolism*
Middle Aged
Neoplasm Invasiveness
PTEN Phosphohydrolase / genetics
PTEN Phosphohydrolase / metabolism*
Proto-Oncogene Proteins c-akt / genetics
Proto-Oncogene Proteins c-akt / metabolism*
RNA Interference
Real-Time Polymerase Chain Reaction
Signal Transduction / drug effects
Tissue Inhibitor of Metalloproteinase-2 / genetics
Tissue Inhibitor of Metalloproteinase-2 / metabolism*
Transcription, Genetic / drug effects
Transfection
Tumor Suppressor Protein p53 / genetics
Tumor Suppressor Protein p53 / metabolism*
Up-Regulation

Substances

Antibiotics, Antineoplastic
CDKN1A protein, human
Cyclin-Dependent Kinase Inhibitor p21
MIRN519 microRNA, human
MicroRNAs
TIMP2 protein, human
TP53 protein, human
Tumor Suppressor Protein p53
Tissue Inhibitor of Metalloproteinase-2
Doxorubicin
AKT3 protein, human
Proto-Oncogene Proteins c-akt
PTEN Phosphohydrolase
PTEN protein, human