Apoptotic speck protein-like, a highly homologous protein to apoptotic speck protein in the pyrin domain, is silenced by DNA methylation and induces apoptosis in human hepatocellular carcinoma

Takahiko Kubo, Junji Yamamoto, Yuko Shikauchi, Yasuharu Niwa, Kenichi Matsubara, Hirohide Yoshikawa

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

We have identified a novel gene encoding a pyrin domain protein of 89 amino acids that is expressed in various tissues including liver, brain, and spleen. The protein is highly homologous to the pyrin domain of apoptosis-associated speck-like protein (ASC). Therefore, we termed it ASC-like (ASCL). We found that ASCL gene was densely and frequently (80%) methylated in hepatocellular carcinoma (HCC) cell lines. In contrast, normal liver samples did not show any significant methylation. This aberrant methylation correlated well with the suppression of RNA expression. Furthermore, a demethylating agent, 5-aza-2′-deoxycytidine, reactivated the ASCL expression in the methylation-silenced cells, indicating that ASCL, is silenced by the associated DNA methylation. ASCL methylation was also found in primary HCC (4 of 17 samples), although the frequency was less than that in cell lines. In addition, we found that ASC was also methylated in primary samples (6 of the 17). Interestingly, either ASCL or ASC methylation was observed in 53% (9 of the 17) of primary HCC samples. Significantly, the restoration of ASCL in the methylation-silenced cells demonstrated growth suppression in colony formation assay. This growth suppression effect of ASCL was supported by apoptotic changes observed in ASCL-transfected cells in which annexin-V binding was positive and caspase-3 was activated. Based on the methylation-silencing and the growth suppression activity, we propose that ASCL plays a significant role in the development of HCC.

Original languageEnglish
Pages (from-to)5172-5177
Number of pages6
JournalCancer Research
Volume64
Issue number15
DOIs
Publication statusPublished - Aug 1 2004

Fingerprint

DNA Methylation
Methylation
Hepatocellular Carcinoma
Apoptosis
Proteins
decitabine
Growth
Cell Line
Annexin A5
Liver
Pyrin Domain
Caspase 3
Genes
Spleen
RNA
Amino Acids
Brain

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Apoptotic speck protein-like, a highly homologous protein to apoptotic speck protein in the pyrin domain, is silenced by DNA methylation and induces apoptosis in human hepatocellular carcinoma. / Kubo, Takahiko; Yamamoto, Junji; Shikauchi, Yuko; Niwa, Yasuharu; Matsubara, Kenichi; Yoshikawa, Hirohide.

In: Cancer Research, Vol. 64, No. 15, 01.08.2004, p. 5172-5177.

Research output: Contribution to journalArticle

@article{9080721dd29048e6b11e3c5daca731f1,
title = "Apoptotic speck protein-like, a highly homologous protein to apoptotic speck protein in the pyrin domain, is silenced by DNA methylation and induces apoptosis in human hepatocellular carcinoma",
abstract = "We have identified a novel gene encoding a pyrin domain protein of 89 amino acids that is expressed in various tissues including liver, brain, and spleen. The protein is highly homologous to the pyrin domain of apoptosis-associated speck-like protein (ASC). Therefore, we termed it ASC-like (ASCL). We found that ASCL gene was densely and frequently (80{\%}) methylated in hepatocellular carcinoma (HCC) cell lines. In contrast, normal liver samples did not show any significant methylation. This aberrant methylation correlated well with the suppression of RNA expression. Furthermore, a demethylating agent, 5-aza-2′-deoxycytidine, reactivated the ASCL expression in the methylation-silenced cells, indicating that ASCL, is silenced by the associated DNA methylation. ASCL methylation was also found in primary HCC (4 of 17 samples), although the frequency was less than that in cell lines. In addition, we found that ASC was also methylated in primary samples (6 of the 17). Interestingly, either ASCL or ASC methylation was observed in 53{\%} (9 of the 17) of primary HCC samples. Significantly, the restoration of ASCL in the methylation-silenced cells demonstrated growth suppression in colony formation assay. This growth suppression effect of ASCL was supported by apoptotic changes observed in ASCL-transfected cells in which annexin-V binding was positive and caspase-3 was activated. Based on the methylation-silencing and the growth suppression activity, we propose that ASCL plays a significant role in the development of HCC.",
author = "Takahiko Kubo and Junji Yamamoto and Yuko Shikauchi and Yasuharu Niwa and Kenichi Matsubara and Hirohide Yoshikawa",
year = "2004",
month = "8",
day = "1",
doi = "10.1158/0008-5472.CAN-03-3314",
language = "English",
volume = "64",
pages = "5172--5177",
journal = "Cancer Research",
issn = "0008-5472",
number = "15",

}

TY - JOUR

T1 - Apoptotic speck protein-like, a highly homologous protein to apoptotic speck protein in the pyrin domain, is silenced by DNA methylation and induces apoptosis in human hepatocellular carcinoma

AU - Kubo, Takahiko

AU - Yamamoto, Junji

AU - Shikauchi, Yuko

AU - Niwa, Yasuharu

AU - Matsubara, Kenichi

AU - Yoshikawa, Hirohide

PY - 2004/8/1

Y1 - 2004/8/1

N2 - We have identified a novel gene encoding a pyrin domain protein of 89 amino acids that is expressed in various tissues including liver, brain, and spleen. The protein is highly homologous to the pyrin domain of apoptosis-associated speck-like protein (ASC). Therefore, we termed it ASC-like (ASCL). We found that ASCL gene was densely and frequently (80%) methylated in hepatocellular carcinoma (HCC) cell lines. In contrast, normal liver samples did not show any significant methylation. This aberrant methylation correlated well with the suppression of RNA expression. Furthermore, a demethylating agent, 5-aza-2′-deoxycytidine, reactivated the ASCL expression in the methylation-silenced cells, indicating that ASCL, is silenced by the associated DNA methylation. ASCL methylation was also found in primary HCC (4 of 17 samples), although the frequency was less than that in cell lines. In addition, we found that ASC was also methylated in primary samples (6 of the 17). Interestingly, either ASCL or ASC methylation was observed in 53% (9 of the 17) of primary HCC samples. Significantly, the restoration of ASCL in the methylation-silenced cells demonstrated growth suppression in colony formation assay. This growth suppression effect of ASCL was supported by apoptotic changes observed in ASCL-transfected cells in which annexin-V binding was positive and caspase-3 was activated. Based on the methylation-silencing and the growth suppression activity, we propose that ASCL plays a significant role in the development of HCC.

AB - We have identified a novel gene encoding a pyrin domain protein of 89 amino acids that is expressed in various tissues including liver, brain, and spleen. The protein is highly homologous to the pyrin domain of apoptosis-associated speck-like protein (ASC). Therefore, we termed it ASC-like (ASCL). We found that ASCL gene was densely and frequently (80%) methylated in hepatocellular carcinoma (HCC) cell lines. In contrast, normal liver samples did not show any significant methylation. This aberrant methylation correlated well with the suppression of RNA expression. Furthermore, a demethylating agent, 5-aza-2′-deoxycytidine, reactivated the ASCL expression in the methylation-silenced cells, indicating that ASCL, is silenced by the associated DNA methylation. ASCL methylation was also found in primary HCC (4 of 17 samples), although the frequency was less than that in cell lines. In addition, we found that ASC was also methylated in primary samples (6 of the 17). Interestingly, either ASCL or ASC methylation was observed in 53% (9 of the 17) of primary HCC samples. Significantly, the restoration of ASCL in the methylation-silenced cells demonstrated growth suppression in colony formation assay. This growth suppression effect of ASCL was supported by apoptotic changes observed in ASCL-transfected cells in which annexin-V binding was positive and caspase-3 was activated. Based on the methylation-silencing and the growth suppression activity, we propose that ASCL plays a significant role in the development of HCC.

UR - http://www.scopus.com/inward/record.url?scp=3442885926&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3442885926&partnerID=8YFLogxK

U2 - 10.1158/0008-5472.CAN-03-3314

DO - 10.1158/0008-5472.CAN-03-3314

M3 - Article

C2 - 15289321

AN - SCOPUS:3442885926

VL - 64

SP - 5172

EP - 5177

JO - Cancer Research

JF - Cancer Research

SN - 0008-5472

IS - 15

ER -