論文 - 時野　隆至

Aberrant methylation of RASGRF1 is associated with an epigenetic field defect and increased risk of gastric cancer.

Takamaru H, Yamamoto E, Suzuki H, Nojima M, Maruyama R, Yamano HO, Yoshikawa K, Kimura T, Harada T, Ashida M, Suzuki R, Yamamoto H, Kai M, Tokino T, Sugai T, Imai K, Toyota M, Shinomura Y

Cancer prevention research (Philadelphia, Pa.)   5 ( 10 ) 1203 - 12  2012年10月  [査読有り]  [国際誌]

　概要を見る

Aberrant DNA methylation is implicated in the epigenetic field defect seen in gastric cancer. Our aim in this study was to identify predictive biomarkers by screening for DNA methylation in noncancerous background gastric mucosa from patients with gastric cancer. Using methylated-CpG island amplification coupled with CpG island microarray (MCAM) analysis, we identified 224 genes that were methylated in the noncancerous gastric mucosa of patients with gastric cancer. Among them, RASGRF1 methylation was significantly elevated in gastric mucosa from patients with either intestinal or diffuse type gastric cancer, as compared with mucosa from healthy individuals (8.3% vs. 22.4%, P < 0.001; 8.3% vs. 19.4%, P < 0.001). RASGRF1 methylation was independent of mucosal atrophy and could be used to distinguish both serum pepsinogen test-positive [sensitivity, 70.0%; specificity, 86.7%; area under the receiver operator characteristic (ROC) curve, AUC, 0.763] and -negative patients with gastric cancer (sensitivity, 72.2%; specificity, 87.0%; AUC, 0.844) from healthy individuals. Ectopic expression of RASGRF1 suppressed colony formation and Matrigel invasion by gastric cancer cells, suggesting it may be involved in gastric tumorigenesis. Collectively, our data suggest that RASGRF1 methylation is significantly involved in an epigenetic field defect in the stomach, and that it could be a useful biomarker to identify individuals at high risk for gastric cancer.

DOI PubMed

DNAメチル化の網羅的解析により同定された新規肝癌関連遺伝子

志谷 真啓, 鈴木 拓, 阿久津 典之, 高木 秀安, 佐々木 茂, 野島 正寛, 山本 博幸, 時野 隆至, 平田 公一, 篠村 恭久

日本消化器病学会雑誌 ( (一財)日本消化器病学会 )  109 ( 臨増大会 ) A700 - A700  2012年09月

ゲノム網羅的shRNAライブラリースクリーニングによるp53誘導アポトーシス阻害遺伝子の同定(The identification of target genes inhibiting p53-induced apoptosis by the screening of genome-wide shRNA library)

大箸 智子, 井戸川 雅史, 佐々木 泰史, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  71回   176 - 176  2012年08月

多発性骨髄腫の発生におけるDNAメチロームの意義 MBDシーケンシング法によるアプローチ(The significance of DNA methylome for tumor progression in multiple myeloma: An MBD-sequencing-based approach)

野島 正寛, 青木 由佳, 安井 寛, 丸山 玲緒, 麻奥 英毅, 石田 禎夫, 時野 隆至, 森 満, 鈴木 拓, 篠村 恭久

日本癌学会総会記事 ( (一社)日本癌学会 )  71回   385 - 385  2012年08月

p53ファミリーの新規標的FOXF1の同定と機能解析 がん細胞の浸潤・遊走能への関与(Forkhead box F1 is a novel target gene of the p53 family and regulates cancer cell migration and invasiveness)

田村 みゆき, 佐々木 泰史, 井戸川 雅史, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  71回   175 - 176  2012年08月

p53ファミリーとがん p53ファミリーネットワークと腫瘍抑制(p53 family and cancer research p53 family network and tumor suppression)

佐々木 泰史, 井戸川 雅史, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  71回   41 - 41  2012年08月

ゲノム網羅的p53応答配列解析とクロマチン免疫沈降シーケンシング(ChIP-seq)を組み合わせた新規p53標的遺伝子の同定(Identification of novel p53 target genes by ChIP-seq combined with genome-wide p53-binding motif analysis in silico)

井戸川 雅史, 大箸 智子, 佐々木 泰史, 篠村 恭久, 今井 浩三, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  71回   257 - 258  2012年08月

CHFR protein regulates mitotic checkpoint by targeting PARP-1 protein for ubiquitination and degradation.

Kashima L, Idogawa M, Mita H, Shitashige M, Yamada T, Ogi K, Suzuki H, Toyota M, Ariga H, Sasaki Y, Tokino T

The Journal of biological chemistry   287 ( 16 ) 12975 - 84  2012年04月  [査読有り]  [国際誌]

　概要を見る

The mitotic checkpoint gene CHFR (checkpoint with forkhead-associated (FHA) and RING finger domains) is silenced by promoter hypermethylation or mutated in various human cancers, suggesting that CHFR is an important tumor suppressor. Recent studies have reported that CHFR functions as an E3 ubiquitin ligase, resulting in the degradation of target proteins. To better understand how CHFR suppresses cell cycle progression and tumorigenesis, we sought to identify CHFR-interacting proteins using affinity purification combined with mass spectrometry. Here we show poly(ADP-ribose) polymerase 1 (PARP-1) to be a novel CHFR-interacting protein. In CHFR-expressing cells, mitotic stress induced the autoPARylation of PARP-1, resulting in an enhanced interaction between CHFR and PARP-1 and an increase in the polyubiquitination/degradation of PARP-1. The decrease in PARP-1 protein levels promoted cell cycle arrest at prophase, supporting that the cells expressing CHFR were resistant to microtubule inhibitors. In contrast, in CHFR-silenced cells, polyubiquitination was not induced in response to mitotic stress. Thus, PARP-1 protein levels did not decrease, and cells progressed into mitosis under mitotic stress, suggesting that CHFR-silenced cancer cells were sensitized to microtubule inhibitors. Furthermore, we found that cells from Chfr knockout mice and CHFR-silenced primary gastric cancer tissues expressed higher levels of PARP-1 protein, strongly supporting our data that the interaction between CHFR and PARP-1 plays an important role in cell cycle regulation and cancer therapeutic strategies. On the basis of our studies, we demonstrate a significant advantage for use of combinational chemotherapy with PARP inhibitors for cancer cells resistant to microtubule inhibitors.

DOI PubMed

Identification of p53 target genes by ChIP-Seq and transcriptome analysis combined with genome-wide p53-binding motif analysis in silico

Masashi Idogawa, Yasushi Sasaki, Yasuhisa Shinomura, Kohzoh Imai, Takashi Tokino

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH )  72  2012年04月  [査読有り]

DOI

p53により発現抑制される分泌性増殖因子HDGFの同定とがん組織における発現解析

佐々木 泰史, 根岸 秀明, 小山 良太, 井戸川 雅史, 鈴木 拓, 今井 浩三, 篠村 恭久, 時野 隆至

日本臨床分子医学会学術総会プログラム・抄録集 ( 日本臨床分子医学会 )  49回   133 - 133  2012年04月

Upregulation of miR-196a and HOTAIR drive malignant character in gastrointestinal stromal tumors.

Niinuma T, Suzuki H, Nojima M, Nosho K, Yamamoto H, Takamaru H, Yamamoto E, Maruyama R, Nobuoka T, Miyazaki Y, Nishida T, Bamba T, Kanda T, Ajioka Y, Taguchi T, Okahara S, Takahashi H, Nishida Y, Hosokawa M, Hasegawa T, Tokino T, Hirata K, Imai K, Toyota M, Shinomura Y

Cancer research   72 ( 5 ) 1126 - 36  2012年03月  [査読有り]  [国際誌]

　概要を見る

Large intergenic noncoding RNAs (lincRNA) have been less studied than miRNAs in cancer, although both offer considerable theranostic potential. In this study, we identified frequent upregulation of miR-196a and lincRNA HOTAIR in high-risk gastrointestinal stromal tumors (GIST). Overexpression of miR-196a was associated with high-risk grade, metastasis and poor survival among GIST specimens. miR-196a genes are located within the HOX gene clusters and microarray expression analysis revealed that the HOXC and HOTAIR gene were also coordinately upregulated in GISTs which overexpress miR-196a. In like manner, overexpression of HOTAIR was also strongly associated with high-risk grade and metastasis among GIST specimens. RNA interference-mediated knockdown of HOTAIR altered the expression of reported HOTAIR target genes and suppressed GIST cell invasiveness. These findings reveal concurrent overexpression of HOX genes with noncoding RNAs in human cancer in this setting, revealing miR-196a and HOTAIR as potentially useful biomarkers and therapeutic targets in malignant GISTs.

DOI PubMed

Genomic vulnerability to LINE-1 hypomethylation is a potential determinant of the clinicogenetic features of multiple myeloma.

Aoki Y, Nojima M, Suzuki H, Yasui H, Maruyama R, Yamamoto E, Ashida M, Itagaki M, Asaoku H, Ikeda H, Hayashi T, Imai K, Mori M, Tokino T, Ishida T, Toyota M, Shinomura Y

Genome medicine   4 ( 12 ) 101 - 101  2012年  [査読有り]  [国際誌]

　概要を見る

BACKGROUND: The aim of this study was to clarify the role of global hypomethylation of repetitive elements in determining the genetic and clinical features of multiple myeloma (MM). METHODS: We assessed global methylation levels using four repetitive elements (long interspersed nuclear element-1 (LINE-1), Alu Ya5, Alu Yb8, and Satellite-α) in clinical samples comprising 74 MM samples and 11 benign control samples (7 cases of monoclonal gammopathy of undetermined significance (MGUS) and 4 samples of normal plasma cells (NPC)). We also evaluated copy-number alterations using array-based comparative genomic hybridization, and performed methyl-CpG binding domain sequencing (MBD-seq). RESULTS: Global levels of the repetitive-element methylation declined with the degree of malignancy of plasma cells (NPC>MGUS>MM), and there was a significant inverse correlation between the degree of genomic loss and the LINE-1 methylation levels. We identified 80 genomic loci as common breakpoints (CBPs) around commonly lost regions, which were significantly associated with increased LINE-1 densities. MBD-seq analysis revealed that average DNA-methylation levels at the CBP loci and relative methylation levels in regions with higher LINE-1 densities also declined during the development of MM. We confirmed that levels of methylation of the 5' untranslated region of respective LINE-1 loci correlated strongly with global LINE-1 methylation levels. Finally, there was a significant association between LINE-1 hypomethylation and poorer overall survival (hazard ratio 2.8, P = 0.015). CONCLUSION: Global hypomethylation of LINE-1 is associated with the progression of and poorer prognosis for MM, possibly due to frequent copy-number loss.

DOI PubMed

2E1412 走査型プローブ顕微鏡を用いた癌抑制因子p53の一分子構造解析(蛋白質-物性II,口頭発表,日本生物物理学会第50回年会(2012年度))

Takahashi Seiya, Sainoo Yasuyuki, Kashima Risa, Tokino Takashi, Komeda Tadahiro, Takahashi Satoshi, Kamagata Kiyoto

生物物理 ( 一般社団法人 日本生物物理学会 )  52   S45 - S46  2012年

DOI CiNii

Erratum

Tomita M, Toyota M, Ishikawa C, Nakazato T, Okudaira T, Matsuda T, Uchihara J. N, Taira N, Ohshiro K, Senba M, Tanaka Y, Ohshima K, Saya H, Tokino T, Mori N

International Journal of Cancer   129 ( 11 ) 2763  2011年12月

DOI

p53 negatively regulates the hepatoma growth factor HDGF.

Sasaki Y, Negishi H, Idogawa M, Yokota I, Koyama R, Kusano M, Suzuki H, Fujita M, Maruyama R, Toyota M, Saito T, Tokino T

Cancer research   71 ( 22 ) 7038 - 47  2011年11月  [査読有り]  [国際誌]

　概要を見る

Hepatoma-derived growth factor (HDGF) is a secreted heparin-binding growth factor that has been implicated in cancer development and progression. Here, we report that HDGF is a critical target for transcriptional repression by the tumor suppressor p53. Endogenous HDGF expression was decreased in cancer cells with introduction of wild-type p53, which also downregulated HDGF expression after DNA damage. In support of the likelihood that HDGF is a critical driver of cancer cell growth, addition of neutralizing HDGF antibodies to culture media was sufficient to block cell growth, migration, and invasion. Similarly, these effects were elicited by conditioned culture medium from p53-expressing cells, and they could be reversed by the addition of recombinant human HDGF. Interestingly, we found that HDGF was overexpressed also in primary gastric, breast, and lung cancer tissues harboring mutant p53 genes. Mechanistic investigations revealed that p53 repressed HDGF transcription by altering HDAC-dependent chromatin remodeling. Taken together, our results reveal a new pathway in which loss of p53 function contributes to the aggressive pathobiological potential of human cancers by elevating HDGF expression.

DOI PubMed

Genome-wide profiling of chromatin signatures reveals epigenetic regulation of MicroRNA genes in colorectal cancer.

Suzuki H, Takatsuka S, Akashi H, Yamamoto E, Nojima M, Maruyama R, Kai M, Yamano HO, Sasaki Y, Tokino T, Shinomura Y, Imai K, Toyota M

Cancer research   71 ( 17 ) 5646 - 58  2011年09月  [査読有り]  [国際誌]

　概要を見る

Altered expression of microRNAs (miRNA) occurs commonly in human cancer, but the mechanisms are generally poorly understood. In this study, we examined the contribution of epigenetic mechanisms to miRNA dysregulation in colorectal cancer by carrying out high-resolution ChIP-seq. Specifically, we conducted genome-wide profiling of trimethylated histone H3 lysine 4 (H3K4me3), trimethylated histone H3 lysine 27 (H3K27me3), and dimethylated histone H3 lysine 79 (H3K79me2) in colorectal cancer cell lines. Combining miRNA expression profiles with chromatin signatures enabled us to predict the active promoters of 233 miRNAs encoded in 174 putative primary transcription units. By then comparing miRNA expression and histone modification before and after DNA demethylation, we identified 47 miRNAs encoded in 37 primary transcription units as potential targets of epigenetic silencing. The promoters of 22 transcription units were associated with CpG islands (CGI), all of which were hypermethylated in colorectal cancer cells. DNA demethylation led to increased H3K4me3 marking at silenced miRNA genes, whereas no restoration of H3K79me2 was detected in CGI-methylated miRNA genes. DNA demethylation also led to upregulation of H3K4me3 and H3K27me3 in a number of CGI-methylated miRNA genes. Among the miRNAs we found to be dysregulated, many of which are implicated in human cancer, miR-1-1 was methylated frequently in early and advanced colorectal cancer in which it may act as a tumor suppressor. Our findings offer insight into the association between chromatin signatures and miRNA dysregulation in cancer, and they also suggest that miRNA reexpression may contribute to the effects of epigenetic therapy.

DOI PubMed

ゲノム網羅的shRNAライブラリースクリーニングに基づく人工miRNA/p53共発現による癌治療(Cancer therapy by the transduction of p53 and artificial miRNAs based on the screening of genome-wide shRNA library)

井戸川 雅史, 佐々木 泰史, 鹿島 理沙, 篠村 恭久, 今井 浩三, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  70回   52 - 52  2011年09月

多発性骨髄腫におけるDNA繰り返し配列の低メチル化と染色体異常の関連(The relationship between DNA hypomethylation of repetitive elements and chromosomal aberrations in multiple myeloma)

青木 由佳, 野島 正寛, 安井 寛, 丸山 玲緒, 鈴木 拓, 石田 禎夫, 麻奥 英毅, 時野 隆至, 豊田 実, 篠村 恭久

日本癌学会総会記事 ( (一社)日本癌学会 )  70回   137 - 137  2011年09月

多発性骨髄腫におけるDNAメチロームと染色体異常 メチル化DNA結合蛋白を利用した次世代シーケンシングによる検討(DNA methylome and chromosomal aberrations in multiple myeloma: an integrated analysis based on MBD-sequencing)

野島 正寛, 青木 由佳, 安井 寛, 丸山 玲緒, 鈴木 拓, 石田 禎夫, 時野 隆至, 篠村 恭久, 豊田 実

日本癌学会総会記事 ( (一社)日本癌学会 )  70回   47 - 47  2011年09月

p53ファミリーの新規標的遺伝子FOXF1の同定と機能解析(Forkhead transcription factor FOXF1 is a novel target gene of the p53 family and regulates cancer cell invasiveness)

田村 みゆき, 佐々木 泰史, 横田 育子, 鹿島 理沙, 井戸川 雅史, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  70回   153 - 153  2011年09月