Identification of epigenetically inactivated genes in human hepatocellular carcinoma by integrative analyses of methylation profiling and pharmacological unmasking.
OBJECTIVE
DNA methylation-dependent transcriptional inactivation of tumor suppressor genes (TSGs) is critical for the pathogenesis of hepatocellular carcinoma (HCC). This study identifies potential TSGs in HCCs using methylation profiling and pharmacological unmasking of methylated TSGs.


METHODS
Methylation profiling was performed on 22 pairs of HCCs and their corresponding noncancerous liver tissues using the Infinium HumanMethylation27 BeadChip. We also determined the gene reexpression after treatment with 5-aza-2'-deoxycytidine (5-Aza-dC) and trichostatin A (TSA) in 5 HCC cell lines.


RESULTS
We selected CpGs that exhibited a significant increase in methylation in HCC tissues compared with that of the noncancerous control group. Two hundred and thirteen CpGs on different gene promoters with a mean difference in the β value ≥0.15 and a value of p < 0.05 were selected. Of the 213 genes, 45 genes were upregulated in 3 or more HCC cell lines with multiplier value of differences ≥2.0 after 5-Aza-dC and TSA treatment.


CONCLUSIONS
We identified several potential TSGs that participate in transcription inactivation through epigenetic interactions in HCC. The results of this study are important for the understanding of functionally important epigenetic alterations in HCC.
Department of Gastroenterology and Hepatology, Kinki University Faculty of Medicine, Osaka-Sayama, Japan.
Naoshi
N
Nishida
Hirokazu
H
Chishina
Tadaaki
T
Arizumi
Masahiro
M
Takita
Satoshi
S
Kitai
Norihisa
N
Yada
Satoru
S
Hagiwara
Tatsuo
T
Inoue
Yasunori
Y
Minami
Kazuomi
K
Ueshima
Toshiharu
T
Sakurai
Masatoshi
M
Kudo
D000368:Aged; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D001707:Biopsy, Needle; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy / Q000235:genetics; D016022:Case-Control Studies; D019175:DNA Methylation; D000077209:Decitabine; D044127:Epigenesis, Genetic; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D020868:Gene Silencing; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D008113:Liver Neoplasms / Q000188:drug therapy / Q000235:genetics; D008297:Male; D008875:Middle Aged; D012016:Reference Values; D012494:Sampling Studies; D014407:Tumor Cells, Cultured / Q000187:drug effects
D006877:Hydroxamic Acids; C012589:trichostatin A; D000077209:Decitabine; D001374:Azacitidine