論文 - 時野　隆至

口腔扁平上皮癌に関与する長鎖非コードRNAの同定

西山 廣陽, 粂川 昴平, 丸山 玲緒, 新沼 猛, 北嶋 洋志, 荻 和弘, 出張 裕也, 甲斐 正広, 宮崎 晃亘, 佐々木 泰史, 時野 隆至, 平塚 博義, 鈴木 拓

日本癌学会総会記事 ( (一社)日本癌学会 )  75回   J - 2018  2016年10月

がん関連遺伝子のエクソーム解析による細胞傷害性抗がん剤感受性マーカーの探索

宇田川 智野, 時野 隆至, 佐々木 泰史, 塩谷 文章, 大西 保行, 末水 洋志, 前佛 均

日本癌学会総会記事 ( (一社)日本癌学会 )  75回   J - 3043  2016年10月

Identification and characterization of the intercellular adhesion molecule-2 gene as a novel p53 target.

Sasaki Y, Tamura M, Takeda K, Ogi K, Nakagaki T, Koyama R, Idogawa M, Hiratsuka H, Tokino T

Oncotarget   7 ( 38 ) 61426 - 61437  2016年09月  [査読有り]  [国際誌]

　概要を見る

The p53 tumor suppressor inhibits cell growth through the activation of both cell cycle arrest and apoptosis, which maintain genome stability and prevent cancer development. Here, we report that intercellular adhesion molecule-2 (ICAM2) is transcriptionally activated by p53. Specifically, ICAM2 is induced by the p53 family and DNA damage in a p53-dependent manner. We identified a p53 binding sequence located within the ICAM2 gene that is responsive to wild-type p53, TAp73, and TAp63. In terms of function, we found that the ectopic expression of ICAM2 inhibited cancer cell migration and invasion. In addition, we demonstrated that silencing endogenous ICAM2 in cancer cells caused a marked increase in extracellular signal-regulated kinase (ERK) phosphorylation levels, suggesting that ICAM2 inhibits migration and invasion of cancer cells by suppressing ERK signaling. Moreover, ICAM2 is underexpressed in human cancer tissues containing mutant p53 as compared to those with wild-type p53. Notably, the decreased expression of ICAM2 is associated with poor survival in patients with various cancers. Our findings demonstrate that ICAM2 induction by p53 has a key role in inhibiting migration and invasion.

DOI PubMed

Plasticity of lung cancer stem-like cells is regulated by the transcription factor HOXA5 that is induced by oxidative stress.

Saijo H, Hirohashi Y, Torigoe T, Horibe R, Takaya A, Murai A, Kubo T, Kajiwara T, Tanaka T, Shionoya Y, Yamamoto E, Maruyama R, Nakatsugawa M, Kanaseki T, Tsukahara T, Tamura Y, Sasaki Y, Tokino T, Suzuki H, Kondo T, Takahashi H, Sato N

Oncotarget   7 ( 31 ) 50043 - 50056  2016年08月  [査読有り]  [国際誌]

　概要を見る

Cancer stem-like cells (CSCs)/cancer-initiating cells (CICs) are reasonable targets for cancer therapy. However, recent studies have revealed that some non-CSCs/CICs have plastic ability and can dedifferentiate into CSCs/CICs. Therefore, an understanding of the molecular mechanisms that control the plasticity is essential to achieve CSC/CIC-targeting therapy. In this study, we analyzed the plasticity of lung cancer cells and found that lung non-CSCs/CICs can dedifferentiate into CSCs/CICs in accordance with the expression of stem cell transcription factor SOX2. SOX2 expression was induced by the transcription factor HOXA5. Oxidative stress repressed the expression of HDAC8 and then induced histone 3 acetylation and increased the expression of HOXA5 and SOX2. These findings indicate that lung cancer cells have plasticity under a condition of oxidative stress and that HOAX5 has a critical role in dedifferentiation.

DOI PubMed

p53 downregulates CRKL oncogene through miR-200

Takashi Tokino, Miyuki Tamura, Masashi Idogawa, Yasushi Sasaki

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH )  76  2016年07月  [査読有り]

DOI

The identification of p53 target genes and noncoding RNAs through the combined analysis of RNA-seq and ChIP-seq data

Masashi Idogawa, Tomoko Ohashi, Yasushi Sasaki, Takashi Tokino

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH )  76  2016年07月  [査読有り]

DOI

Semiconductor-based next-generation sequencing analysis of 409 cancer-related genes for mutations and copy-number variations in oral squamous cell carcinoma

Takafumi Nakagaki, Yasushi Sasaki, Masashi Idogawa, Ryota Koyama, Kenta Kobashi, Miyuki Tamura, Tomoko Ohashi, Kazuhiro Ogi, Hiroyoshi Hiratsuka, Takashi Tokino

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH )  76  2016年07月  [査読有り]

DOI

Targeted sequencing of 409 cancer related genes for somatic mutations and copy number variations in human cancer using the semiconductor sequencers

Yasushi Sasaki, Ryota Koyama, Takafumi Nakagaki, Miyuki Tamura, Masashi Idogawa, Takashi Tokino

CANCER RESEARCH ( AMER ASSOC CANCER RESEARCH )  76  2016年07月  [査読有り]

DOI

A genomic screen for long noncoding RNA genes epigenetically silenced by aberrant DNA methylation in colorectal cancer.

Kumegawa K, Maruyama R, Yamamoto E, Ashida M, Kitajima H, Tsuyada A, Niinuma T, Kai M, Yamano HO, Sugai T, Tokino T, Shinomura Y, Imai K, Suzuki H

Scientific reports   6   26699 - 26699  2016年05月  [査読有り]  [国際誌]

　概要を見る

Long noncoding RNAs (lncRNAs) have emerged as key components in multiple cellular processes, although their physiological and pathological functions are not fully understood. To identify cancer-related lncRNAs, we screened for those that are epigenetically silenced in colorectal cancer (CRC). Through a genome-wide analysis of histone modifications in CRC cells, we found that the transcription start sites (TSSs) of 1,027 lncRNA genes acquired trimethylation of histone H3 lysine 4 (H3K4me3) after DNA demethylation. Integrative analysis of chromatin signatures and the DNA methylome revealed that the promoter CpG islands (CGIs) of 66 lncRNA genes contained cancer-specific methylation. By validating the expression and methylation of lncRNA genes in CRC cells, we ultimately identified 20 lncRNAs, including ZNF582-AS1, as targets of epigenetic silencing in CRC. ZNF582-AS1 is frequently methylated in CRC cell lines (87.5%), primary CRCs (77.2%), colorectal adenomas (44.7%) and advanced adenomas (87.8%), suggesting that this methylation is an early event during colorectal tumorigenesis. Methylation of ZNF582-AS1 is associated with poor survival of CRC patients, and ectopic expression of ZNF582-AS1 suppressed colony formation by CRC cells. Our findings offer insight into the association between epigenetic alterations and lncRNA dysregulation in cancer and suggest that ZNF582-AS1 may be a novel tumor-suppressive lncRNA.

DOI PubMed

Low-grade mucoepidermoid carcinoma with regional lymph node metastasis: A case report and genetic review of criteria for grading

Takafumi Nakagaki, Kazuhiro Ogi, Masato Abe, Hironari Dehari, Akihiro Miyazaki, Tadashi Hasegawa, Takashi Tokino, Hiroyoshi Hiratsuka

Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology ( Elsevier Ltd )  28 ( 2 ) 189 - 192  2016年03月  [査読有り]

　概要を見る

Mucoepidermoid carcinoma (MEC) is the most common salivary gland malignancy. It accounts for 5% of all salivary gland tumors and 20% of salivary malignancies and has various clinical outcomes. Its prognosis is related to the tumor grade in the WHO classification of head and neck tumors. MEC is divided into high, intermediate and low grades. In general, low-grade MEC with cervical lymph node metastasis is rare, and primary MEC that occurs in a minor salivary gland with cervical lymph node metastasis is very rare. Low-grade lesions have quite different behavior from high-grade ones, leading to differences in the treatment course required for disease control. MEC is associated with recurring chromosomal translocation t(11 19)(q21 p13). CRTC1-MAML2 gene fusion was identified from the recurring translocation, which is often the sole cytogenetic alteration in this disease. This fusion transcript is detected frequently in MEC and is involved in the transformation of epithelial cells. Detection of the CRTC1-MAML2 fusion transcript is associated with a less advanced clinical stage and low-grade tumor histology. The presence of the transcript is associated with longer disease-free and overall survival periods.Here, we report a rare case of low-grade MEC with cervical lymph node metastasis. The findings for this patient show that the CRTC1-MAML2 fusion transcript may be specific to MEC, and associated with a distinct MEC subset that exhibits favorable clinicopathologic features.

DOI

Genomic characterization of esophageal squamous cell carcinoma: Insights from next-generation sequencing.

Sasaki Y, Tamura M, Koyama R, Nakagaki T, Adachi Y, Tokino T

World journal of gastroenterology   22 ( 7 ) 2284 - 93  2016年02月  [査読有り]  [国際誌]

　概要を見る

Two major types of cancer occur in the esophagus: squamous cell carcinoma, which is associated with chronic smoking and alcohol consumption, and adenocarcinoma, which typically arises in gastric reflux-associated Barrett's esophagus. Although there is increasing incidence of esophageal adenocarcinoma in Western counties, esophageal squamous cell carcinoma (ESCC) accounts for most esophageal malignancies in East Asia, including China and Japan. Technological advances allowing for massively parallel, high-throughput next-generation sequencing (NGS) of DNA have enabled comprehensive characterization of somatic mutations in large numbers of tumor samples. Recently, several studies were published in which whole exome or whole genome sequencing was performed in ESCC tumors and compared with matched normal DNA. Mutations were validated in several genes, including in TP53, CDKN2A, FAT1, NOTCH1, PIK3CA, KMT2D and NFE2L2, which had been previously implicated in ESCC. Several new recurrent alterations have also been identified in ESCC. Combining the clinicopathological characteristics of patients with information obtained from NGS studies may lead to the development of effective diagnostic and therapeutic approaches for ESCC. As this research becomes more prominent, it is important that gastroenterologist become familiar with the various NGS technologies and the results generated using these methods. In the present study, we describe recent research approaches using NGS in ESCC.

DOI PubMed

TET1 Depletion Induces Aberrant CpG Methylation in Colorectal Cancer Cells.

Masahiro Kai, Takeshi Niinuma, Hiroshi Kitajima, Eiichiro Yamamoto, Taku Harada, Hironori Aoki, Reo Maruyama, Mutsumi Toyota, Yasushi Sasaki, Tamotsu Sugai, Takashi Tokino, Hiroshi Nakase, Hiromu Suzuki

PloS one   11 ( 12 ) e0168281  2016年  [査読有り]  [国際誌]

　概要を見る

Aberrant DNA methylation is commonly observed in colorectal cancer (CRC), but the underlying mechanism is not fully understood. 5-hydroxymethylcytosine levels and TET1 expression are both reduced in CRC, while epigenetic silencing of TET1 is reportedly associated with the CpG island methylator phenotype. In the present study, we aimed to clarify the relationship between loss of TET1 and aberrant DNA methylation in CRC. Stable TET1 knockdown clones were established using Colo320DM cells, which express high levels of TET1, and HCT116 cells, which express TET1 at a level similar to that in normal colonic tissue. Infinium HumanMethylation450 BeadChip assays revealed increased levels of 5-methylcytosine at more than 10,000 CpG sites in TET1-depleted Colo320DM cells. Changes in DNA methylation were observed at various positions within the genome, including promoters, gene bodies and intergenic regions, and the altered methylation affected expression of a subset of genes. By contrast, TET1 knockdown did not significantly affect DNA methylation in HCT116 cells. However, TET1 depletion was associated with attenuated effects of 5-aza-2'-deoxycytidine on gene expression profiles in both cell lines. These results suggest that loss of TET1 may induce aberrant DNA methylation and may attenuate the effect of 5-aza-2'-deoxycytidine in CRC cells.

DOI PubMed

癌細胞におけるゲノムワイドRNAiスクリーニングを用いたp53誘導アポトーシス抵抗性を解除するshRNAの探索

梅田 璃子, 井戸川 雅史, 大箸 智子, 佐々木 泰史, 時野 隆至

日本生化学会大会・日本分子生物学会年会合同大会講演要旨集 ( (公社)日本生化学会 )  88回・38回   [2LBA040] - [2LBA040]  2015年12月

新規p53標的遺伝子LIMA1の発現誘導は癌細胞浸潤の抑制に必要である

大箸 智子, 井戸川 雅史, 梅田 璃子, 佐々木 泰史, 時野 隆至

日本生化学会大会・日本分子生物学会年会合同大会講演要旨集 ( (公社)日本生化学会 )  88回・38回   [4T18 - 03(3P1065)]  2015年12月

次世代シーケンサーを用いたがん関連409遺伝子のターゲットシーケンス解析

佐々木 泰史, 小山 良太, 中垣 貴文, 田村 みゆき, 大箸 智子, 井戸川 雅史, 時野 隆至

日本生化学会大会・日本分子生物学会年会合同大会講演要旨集 ( (公社)日本生化学会 )  88回・38回   [3LBA004] - [3LBA004]  2015年12月

Epigenetic silencing of NTSR1 is associated with lateral and noninvasive growth of colorectal tumors.

Kamimae S, Yamamoto E, Kai M, Niinuma T, Yamano HO, Nojima M, Yoshikawa K, Kimura T, Takagi R, Harada E, Harada T, Maruyama R, Sasaki Y, Tokino T, Shinomura Y, Sugai T, Imai K, Suzuki H

Oncotarget   6 ( 30 ) 29975 - 90  2015年10月  [査読有り]  [国際誌]

　概要を見る

Our aim was to identify DNA methylation changes associated with the growth pattern and invasiveness of colorectal cancers (CRCs). Comparison of the methylation statuses of large (≥ 20 mm in diameter along the colonic surface) noninvasive tumors (NTs) and small (<20 mm in diameter along the colonic surface) invasive tumors (ITs) using CpG island microarray analysis showed neurotensin receptor 1 (NTSR1) to be hypermethylated in large NTs. Quantitative bisulfite pyrosequencing revealed that NTSR1 is frequently methylated in colorectal tumors, with large NTs exhibiting the highest methylation levels. The higher NTSR1 methylation levels were associated with better prognoses. By contrast, NTSR1 copy number gains were most frequent among small ITs. Methylation of NTSR1 was associated with the gene's silencing in CRC cell lines, whereas ectopic expression of NTSR1 promoted proliferation and invasion by CRC cells. Analysis of primary tumors composed of adenomatous and malignant portions revealed that NTSR1 is frequently methylated in the adenomatous portion, while methylation levels are generally lower in the cancerous portions. These results suggest that NTSR1 methylation is associated with lateral and noninvasive growth of colorectal tumors, while low levels of methylation may contribute to the malignant potential through activation of NTSR1. Our data also indicate that NTSR1 methylation may be a prognostic biomarker in CRC.

DOI PubMed

半導体シーケンサーを用いたがん関連遺伝子のターゲットシーケンス解析

佐々木 泰史, 中垣 貴文, 田村 みゆき, 竹田 康佑, 大箸 智子, 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 )  74回   P - 1156  2015年10月

p53による癌細胞浸潤の抑制には新規標的遺伝子LIMA1の発現誘導が必要である

大箸 智子, 井戸川 雅史, 田村 みゆき, 竹田 康佑, 中垣 貴文, 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 )  74回   J - 1280  2015年10月

CRKLはmiR-200ファミリーを介してp53により発現抑制される

田村 みゆき, 佐々木 泰史, 竹田 康佑, 中垣 貴文, 大箸 智子, 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 )  74回   P - 3013  2015年10月

次世代半導体シーケンサーを用いた口腔扁平上皮癌におけるがん関連遺伝子の網羅的解析

中垣 貴文, 佐々木 泰史, 井戸川 雅史, 竹田 康佑, 田村 みゆき, 大箸 智子, 荻 和弘, 平塚 博義, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  74回   J - 1330  2015年10月