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Deregulation of RB1 expression by loss of imprinting in human hepatocellular carcinoma
Author(s) -
Anwar Sumadi Lukman,
Krech Till,
Hasemeier Britta,
Schipper Elisa,
Schweitzer Nora,
Vogel Arndt,
Kreipe Hans,
Lehmann Ulrich
Publication year - 2014
Publication title -
the journal of pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.964
H-Index - 184
eISSN - 1096-9896
pISSN - 0022-3417
DOI - 10.1002/path.4376
Subject(s) - dna methylation , biology , methylation , cpg site , loss of heterozygosity , hepatocellular carcinoma , locus (genetics) , intron , microbiology and biotechnology , imprinting (psychology) , genomic imprinting , cancer research , gene , gene expression , genetics , allele
The tumour suppressor gene RB 1 is frequently silenced in many different types of human cancer, including hepatocellular carcinoma (HCC). However, mutations of the RB1 gene are relatively rare in HCC . A systematic screen for the identification of imprinted genes deregulated in human HCC revealed that RB1 shows imprint abnormalities in a high proportion of primary patient samples. Altogether, 40% of the HCC specimens (16/40) showed hyper‐ or hypomethylation at the CpG island in intron 2 of the RB1 gene. Re‐analysis of publicly available genome‐wide DNA methylation data confirmed these findings in two independent HCC cohorts. Loss of correct DNA methylation patterns at the RB1 locus leads to the aberrant expression of an alternative RB1–E2B transcript, as measured by quantitative real‐time PCR. Demethylation at the intron 2 CpG island by DNMT1 knock‐down or aza‐deoxycytidine (DAC) treatment stimulated expression of the RB1–E2B transcript, accompanied by diminished RB1 main transcript expression. No aberrant DNA methylation was found at the RB1 locus in hepatocellular adenoma (HCA, n = 10), focal nodular hyperplasia (FNH, n = 5) and their corresponding adjacent liver tissue specimens. Deregulated RB1 expression due to hyper‐ or hypomethylation in intron 2 of the RB1 gene is found in tumours without loss of heterozygosity and is associated with a decrease in overall survival ( p = 0.032) if caused by hypermethylation of CpG85. This unequivocally demonstrates that loss of imprinting represents an important additional mechanism for RB1 pathway inactivation in human HCC , complementing well‐described molecular defects. Copyright © 2014 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.