論文 - 時野　隆至

新規p53標的遺伝子として同定されたp120カテニンファミリー蛋白の癌における役割(The roles of p120 catenin family protein as a novel p53 target in cancer)

鈴木 菜摘, 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 )  77回   1301 - 1301  2018年09月

日本人口腔扁平上皮癌における癌関連50遺伝子を標的とした次世代シークエンサーによる解析(Targeted next-generation sequencing of 50 cancer-related genes in Japanese patients with oral squamous cell carcinoma)

荻 和弘, 中垣 貴文, 井戸川 雅史, 宮崎 晃亘, 時野 隆至, 佐々木 泰史

日本癌学会総会記事 ( (一社)日本癌学会 )  77回   784 - 784  2018年09月

大腸癌における発現ネットワーク解析によるハブ長鎖非コードRNA(hub-lncRNA)の同定と機能解析(Identification of hub-long non-coding RNAs(lncRNAs) by the network analysis of lncRNA expression in colorectal cancers)

井戸川 雅史, 鈴木 菜摘, 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 )  77回   463 - 463  2018年09月

食道癌肉腫の遺伝子変異解析(Exome sequence analysis of carcinomatous and sarcomatous elements of an esophageal carcinosarcoma)

佐々木 教之, 岩谷 岳, 遠藤 史隆, 鴻巣 正史, 秋山 有史, 佐々木 泰史, 時野 隆至, 西塚 哲

日本癌学会総会記事 ( (一社)日本癌学会 )  77回   326 - 326  2018年09月

大腸がんにおける腫瘍血管内皮関連遺伝子の同定(Identification and characterization of a tumor endothelium-related gene in colorectal cancer)

萬 顕, 山本 英一郎, 沼田 有斗, 新沼 猛, 北嶋 洋志, 甲斐 正広, 須藤 豪太, 黒瀬 誠, 時野 隆至, 仲瀬 裕志, 菅井 有, 鈴木 拓

日本癌学会総会記事 ( 日本癌学会 )  77回   1574 - 1574  2018年09月

Mutational analysis of uterine cervical cancer that survived multiple rounds of radiotherapy.

Endang Nuryadi, Yasushi Sasaki, Yoshihiko Hagiwara, Tiara Bunga Mayang Permata, Hiro Sato, Shuichiro Komatsu, Yuya Yoshimoto, Kazutoshi Murata, Ken Ando, Nobuteru Kubo, Noriyuki Okonogi, Yosuke Takakusagi, Akiko Adachi, Mototaro Iwanaga, Keisuke Tsuchida, Tomoaki Tamaki, Shin-Ei Noda, Yuka Hirota, Atsushi Shibata, Tatsuya Ohno, Takashi Tokino, Takahiro Oike, Takashi Nakano

Oncotarget   9 ( 66 ) 32642 - 32652  2018年08月  [査読有り]  [国際誌]

　概要を見る

Radiotherapy is an essential component of cancer therapy. Despite advances in cancer genomics, the mutation signatures of radioresistant tumors have not yet been fully elucidated. To address this issue, we analyzed a unique set of clinical specimens from a uterine cervical cancer that repeatedly locally recurred after multiple rounds of radiotherapy. Exon sequencing of 409 cancer-related genes in the treatment-naïve tumor and the tumors that recurred after initial and secondary radiotherapy identified (i) activating mutations in PIK3CA and KRAS, and putative inactivating mutations in SMAD4, as trunk mutation signatures that persisted over the clinical course; and (ii) mutations in KMT2A, TET1, and NLRP1 as acquired mutation signatures observed only in recurrent tumors after radiotherapy. Comprehensive mining of published in vitro genomics data pertaining to radiosensitivity revealed that simultaneous mutations in KRAS and SMAD4, which have not been described previously in uterine cervical cancer, are associated with cancer cell radioresistance. The association between this mutation signature and radioresistance was validated by isogenic cell-based experiments. These results provide proof-of-principle for the analytical pipeline employed in this study, which explores clinically relevant mutation signatures for radioresistance, and demonstrate that this approach is worth pursuing with larger cohorts in the future.

DOI PubMed

Screening for long noncoding RNAs associated with oral squamous cell carcinoma reveals the potentially oncogenic actions of DLEU1.

Nishiyama K, Maruyama R, Niinuma T, Kai M, Kitajima H, Toyota M, Hatanaka Y, Igarashi T, Kobayashi JI, Ogi K, Dehari H, Miyazaki A, Yorozu A, Yamamoto E, Idogawa M, Sasaki Y, Sugai T, Tokino T, Hiratsuka H, Suzuki H

Cell death & disease   9 ( 8 ) 826 - 826  2018年08月  [査読有り]  [国際誌]

　概要を見る

Recent studies have shown that long noncoding RNAs (lncRNAs) have pivotal roles in human malignancies, although their significance in oral squamous cell carcinoma (OSCC) is not fully understood. In the present study, we identified lncRNAs functionally associated with OSCC. By analyzing RNA-seq datasets obtained from primary head and neck squamous cell carcinoma (HNSCC), we identified 15 lncRNAs aberrantly expressed in cancer tissues. We then validated their expression in 18 OSCC cell lines using qRT-PCR and identified 6 lncRNAs frequently overexpressed in OSCC. Among those, we found that knocking down DLEU1 (deleted in lymphocytic leukemia 1) strongly suppressed OSCC cell proliferation. DLEU1 knockdown also suppressed migration, invasion, and xenograft formation by OSCC cells, which is suggestive of its oncogenic functionality. Microarray analysis revealed that DLEU1 knockdown significantly affects expression of a number of cancer-related genes in OSCC cells, including HAS3, CD44, and TP63, suggesting that DLEU1 regulates HA-CD44 signaling. Expression of DLEU1 was elevated in 71% of primary OSCC tissues, and high DLEU1 expression was associated with shorter overall survival of HNSCC patients. These data suggest that elevated DLEU1 expression contributes to OSCC development, and that DLEU1 may be a useful therapeutic target in OSCC.

DOI PubMed

Target amplicon exome-sequencing identifies promising diagnosis and prognostic markers involved in RTK-RAS and PI3K-AKT signaling as central oncopathways in primary central nervous system lymphoma.

Takashima Y, Sasaki Y, Hayano A, Homma J, Fukai J, Iwadate Y, Kajiwara K, Ishizawa S, Hondoh H, Tokino T, Yamanaka R

Oncotarget   9 ( 44 ) 27471 - 27486  2018年06月  [査読有り]  [国際誌]

　概要を見る

Exome-sequencing for somatic mutation detection and copy number variation analysis are effective and valid methods for evaluating human cancers in current molecular medicine. We conducted target amplicon exome-sequencing analyses using PCR target enrichment and next-generation sequencing on Ion Proton semiconductor sequencers. Twenty-seven primary central nervous system lymphoma (PCNSL) specimens and their corresponding noncancerous tissues were used for multiplex PCR amplification to obtain targeted coverages of the entire coding regions of 409 cancer-related genes. The average of the total numbers of somatic mutations including single-nucleotide variations and insertion/deletion mutations in each specimen was 13.3. Of these, the average of the ratios of nonsynonymous substitutions in each specimen was 74.8%. The most frequent mutations in 27 specimens were in PIM1, MYD88, CD79B, DST, IRF4, ERBB3, MYH11, DCC, and KMT2D. Furthermore, somatic mutations of MYH11 were related to poor prognoses in PCNSL patients. Copy number variations were also duplicated and/or deleted from deep-sequencing in segmental genomic islands. In addition to these prognostic marker candidates, analysis of RTK-RAS-MAPK signaling and the PTEN-PI3K-AKT proapoptotic pathway showed that somatic activations and aberrations, respectively, may be involved in a promising central oncopathway harboring mTOR, c-Myc, FOXO1, and p53. This study provides a foundation for molecular targeted therapies based on genome diagnostics and prognosis in PCNSL.

DOI PubMed

Epigenetic silencing of miR-200b is associated with cisplatin resistance in bladder cancer.

Shindo T, Niinuma T, Nishiyama N, Shinkai N, Kitajima H, Kai M, Maruyama R, Tokino T, Masumori N, Suzuki H

Oncotarget   9 ( 36 ) 24457 - 24469  2018年05月  [査読有り]  [国際誌]

　概要を見る

In this study, we identified microRNAs (miRNAs) involved in cisplatin (CDDP) resistance in bladder cancer (BCa). After establishing CDDP-resistant BCa cell lines (T24RC and EJ138RC), TaqMan arrays revealed that members of the miR-200 family (miR-200b, miR-200a and miR-429) were downregulated in T24RC as compared to parental T24 cells. miR-200b was associated with CDDP sensitivity in BCa cells, and its downregulation was associated with CpG island hypermethylation. Pharmacological demethylation using 5-aza-2'-deoxycytidine restored miR-200b expression, and the combination of 5-aza-2'-deoxycytidine + CDDP strongly inhibited T24RC cell proliferation. Microarray analysis revealed that miR-200b + CDDP induced genes involved in CDDP sensitivity or cytotoxicity, including IGFBP3, ICAM1 and TNFSF10, in the resistant cells. Expression and DNA methylation of miR-200b were inversely associated in primary BCa, and low expression/high methylation was associated with poor overall survival. These results suggest downregulation of miR-200b is associated with CDDP resistance in BCa. Epigenetic silencing of miR-200b may be a marker of CDDP resistance and a useful therapeutic target for overcoming CDDP resistance in BCa.

DOI PubMed

Targeted sequencing reveals genetic variants associated with sensitivity of 79 human cancer xenografts to anticancer drugs.

Udagawa C, Sasaki Y, Suemizu H, Ohnishi Y, Ohnishi H, Tokino T, Zembutsu H

Experimental and therapeutic medicine   15 ( 2 ) 1339 - 1359  2018年02月  [査読有り]  [国際誌]

　概要を見る

Although there has been progress moving from a 'one-size-fits-all' cytotoxic approach to personalized molecular medicine, the majority of patients with cancer receive chemotherapy using cytotoxic anticancer drugs. The sequencing analysis of 409 genes associated with cancer was conducted in the present study using 59 DNA sequences extracted from human cancer xenografts implanted into nude mice, of which sensitivity to 9 cytotoxic anticancer drugs [5-fluorouracil, nimustine, adriamycin, cyclophosphamide, cisplatin, mitomycin C (MMC), methotrexate, vincristine (VCR), and vinblastine] was examined. The present study investigated the association between the sensitivities of the xenografts to the 9 anticancer drugs and the frequency of single nucleotide variants (SNV). The correlation between the expression level of the genes and sensitivities to the 9 drugs in the above xenografts was also estimated. In the screening study using 59 xenografts, 3 SNVs (rs1805321, rs62456182 in PMS1 Homolog 2, Mismatch Repair System Component and rs13382825 in LDL Receptor Related Protein 1B), were associated with sensitivity to VCR and MMC, respectively (P<0.001). A replication study of 596 SNVs was subsequently performed, which indicated P<0.05 in the screening study using independent samples of 20 xenografts. A combined result of the screening and replication studies indicated that 35 SNVs were potentially associated with sensitivities to one or more of the nine anticancer drugs (Pcombined=0.0011-0.035). Of the 35 SNVs, rs16903989 and rs201432181 in Leukemia Inhibitory Factor Receptor α and Adhesion G Protein-Coupled Receptor A2 were commonly associated with sensitivity to 2 or 4 anticancer drugs, respectively. These findings provide novel insights which may benefit the development of personalized anticancer therapy for patients with cancer in the future.

DOI PubMed

Profiling cancer-related gene mutations in oral squamous cell carcinoma from Japanese patients by targeted sequencing

Nakagaki Takafumi, Sasaki Yasushi, Idogawa Masashi, Koyama Ryouta, Tamura Miyuki, Fukushima Hisayo, Ogi Kazuhiro, Hiratsuka Hiroyoshi, Tokino Takashi

CANCER SCIENCE   109   57  2018年01月  [査読有り]

Exploitation of a liquid biopsy system using circulating tumor DNA in esophageal cancer patients.

Iwaya Takeshi, Endo Fuinitaka, Sasaki Yasushi, Yaegashi Mizunori, Chiba Takehiro, Akiyama Yuji, Masuda Mar, Yamada Tesshi, Tokino Takashi, Nishizuka Satoshi

CANCER SCIENCE   109   953  2018年01月  [査読有り]

Identification of genomic alterations in oral squamous cell carcinomas using amplicon-based whole-exome sequencing

Sasaki Yasushi, Tamura Miyuki, Nakagaki Takafumi, Koyama Ryota, Idogawa Masashi, Ogi Kazuhiro, Hiratsuka Hiroyoshi, Nakase Hiroshi, Tokino Takashi

CANCER SCIENCE   109   990  2018年01月  [査読有り]

Exploration of p53-targeting long non-coding RNAs (lncRNAs) and network analysis of lncRNA expression in cancers

Idogawa Masashi, Ohashi Tomoko, Sasaki Yasushi, Tokino Takashi

CANCER SCIENCE   109   82  2018年01月  [査読有り]

Identification and characterization of a metastatic suppressor BRMS1L as a target gene of p53.

Koyama R, Tamura M, Nakagaki T, Ohashi T, Idogawa M, Suzuki H, Tokino T, Sasaki Y

Cancer science   108 ( 12 ) 2413 - 2421  2017年12月  [査読有り]  [国際誌]

　概要を見る

The tumor suppressor p53 and its family members, p63 and p73, play a pivotal role in the cell fate determination in response to diverse upstream signals. As transcription factors, p53 family proteins regulate a number of genes that are involved in cell cycle arrest, apoptosis, senescence, and maintenance of genomic stability. Recent studies revealed that p53 family proteins are important for the regulation of cell invasion and migration. Microarray analysis showed that breast cancer metastasis suppressor 1-like (BRMS1L) is upregulated by p53 family proteins, specifically p53, TAp63γ, and TAp73β. We identified two responsive elements of p53 family proteins in the first intron and upstream of BRMS1L. These response elements are well conserved among mammals. Functional analysis showed that ectopic expression of BRMS1L inhibited cancer cell invasion and migration; knockdown of BRMS1L by siRNA induced the opposite effect. Importantly, clinical databases revealed that reduced BRMS1L expression correlated with poor prognosis in patients with breast and brain cancer. Together, these results strongly indicate that BRMS1L is one of the mediators downstream of the p53 pathway, and that it inhibits cancer cell invasion and migration, which are essential steps in cancer metastasis. Collectively, our results indicate that BRMS1L is involved in cancer cell invasion and migration, and could be a therapeutic target for cancer.

DOI PubMed

Identification and characterization of a metastatic suppressor BRMS1L as a target gene of p53

Ryota Koyama, Miyuki Tamura, Takafumi Nakagaki, Tomoko Ohashi, Masashi Idogawa, Hiromu Suzuki, Takashi Tokino, Yasushi Sasaki

CANCER SCIENCE ( WILEY )  108 ( 12 ) 2413 - 2421  2017年12月  [査読有り]

　概要を見る

The tumor suppressor p53 and its family members, p63 and p73, play a pivotal role in the cell fate determination in response to diverse upstream signals. As transcription factors, p53 family proteins regulate a number of genes that are involved in cell cycle arrest, apoptosis, senescence, and maintenance of genomic stability. Recent studies revealed that p53 family proteins are important for the regulation of cell invasion and migration. Microarray analysis showed that breast cancer metastasis suppressor 1-like (BRMS1L) is upregulated by p53 family proteins, specifically p53, TAp63, and TAp73. We identified two responsive elements of p53 family proteins in the first intron and upstream of BRMS1L. These response elements are well conserved among mammals. Functional analysis showed that ectopic expression of BRMS1L inhibited cancer cell invasion and migration; knockdown of BRMS1L by siRNA induced the opposite effect. Importantly, clinical databases revealed that reduced BRMS1L expression correlated with poor prognosis in patients with breast and brain cancer. Together, these results strongly indicate that BRMS1L is one of the mediators downstream of the p53 pathway, and that it inhibits cancer cell invasion and migration, which are essential steps in cancer metastasis. Collectively, our results indicate that BRMS1L is involved in cancer cell invasion and migration, and could be a therapeutic target for cancer.

DOI PubMed

p53標的遺伝子BRMS1Lの同定と機能解析

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

生命科学系学会合同年次大会 ( 生命科学系学会合同年次大会運営事務局 )  2017年度   [1LBA - 057]  2017年12月

新規p53標的である蛋白コード遺伝子LIMA1と長鎖非コードRNA(IncRNA)NEAT1のp53機能における役割

鈴木 菜摘, 井戸川 雅史, 大箸 智子, 佐々木 泰史, 時野 隆至

生命科学系学会合同年次大会 ( 生命科学系学会合同年次大会運営事務局 )  2017年度   [1P - 0990]  2017年12月

Downregulation of miR-186 is associated with metastatic recurrence of gastrointestinal stromal tumors.

Takeshi Niinuma, Masahiro Kai, Hiroshi Kitajima, Eiichiro Yamamoto, Taku Harada, Reo Maruyama, Takayuki Nobuoka, Toshirou Nishida, Tatsuo Kanda, Tadashi Hasegawa, Takashi Tokino, Tamotsu Sugai, Yasuhisa Shinomura, Hiroshi Nakase, Hiromu Suzuki

Oncology letters   14 ( 5 ) 5703 - 5710  2017年11月  [査読有り]  [国際誌]

　概要を見る

Although dysregulation of microRNAs (miRNAs/miRs) is a common feature of human malignancies, its involvement in gastrointestinal stromal tumors (GISTs) is not fully understood. The present study aimed to identify the miRNAs that perform a role in GIST metastasis. miRNA expression profiles from a series of 32 primary GISTs were analyzed using microarrays, and miR-186 was observed to be downregulated in tumors exhibiting metastatic recurrence. Reverse transcription-quantitative polymerase chain reaction analysis of an independent cohort of 100 primary GISTs revealed that low miR-186 expression is associated with metastatic recurrence and a poor prognosis. Inhibition of miR-186 in GIST-T1 cells promoted cell migration. Gene expression microarray analysis demonstrated that miR-186 inhibition upregulated a set of genes implicated in cancer metastasis, including insulin-like growth factor-binding protein 3, AKT serine/threonine kinase 2, hepatocyte growth factor receptor, CXC chemokine receptor 4 and epidermal growth factor-containing fibulin-like extracellular matrix protein 1. These results suggest that the downregulation of miR-186 is involved in the metastatic recurrence of GISTs, and that miR-186 levels could potentially be a predictive biomarker for clinical outcome.

DOI PubMed

口腔扁平上皮癌の全エクソン解析

佐々木 泰史, 田村 みゆき, 中垣 貴文, 小山 良太, 井戸川 雅史, 荻 和弘, 平塚 博義, 仲瀬 裕志, 時野 隆至

日本癌学会総会記事 ( (一社)日本癌学会 )  76回   P - 3044  2017年09月