研究者詳細 - 佐藤　しのぶ

論文 - 佐藤　しのぶ

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Cyclic Naphthalene Diimide Dimer with a Strengthened Ability to Stabilize Dimeric G-Quadruplex 査読有り国際誌

Takeuchi R., Zou T., Wakahara D., Nakano Y., Sato S., Takenaka S.

Chemistry - A European Journal 25 ( 37 ) 8691 - 8695 2019年07月

記述言語：英語掲載種別：研究論文（学術雑誌）

© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim A new type of dimeric cyclic naphthalene diimide derivatives (cNDI-dimers) carrying varied linker length were designed and synthesized to recognize dimeric G-quadruplex structures. All of the cNDI-dimers exhibited a high preference for recognizing G-quadruplex structures, and significantly enhanced the thermal stability of the dimeric G-quadruplex structure over the cNDI monomer by increasing the melting temperature by more than 23 °C, which indicated the strengthened ability of cNDI dimers for stabilizing dimeric G-quadruplex. cNDI dimers also showed a stronger ability to inhibit telomerase activity and stop telomere DNA elongation than cNDI monomer, which showed an improved anticancer potentiality for further therapeutic application.

DOI： 10.1002/chem.201901468

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Electrochemical Aberrant Methylation Detection Based on Ferrocenyl Naphthalene Diimide Carrying β-cyclodextrin, FNC 招待有り査読有り国際誌

Sato S., Nishi Y., Takenaka S.

Electroanalysis 31 ( 10 ) 1988 - 1993 2019年01月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）

© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Electrochemical detection of the specific gene carrying aberrant methylated cytosine was achieved by ferrocenyl naphthalene diimide carrying β-cyclodextrin (β-CD), FNC, coupled with the probe-DNA-immobilized electrode. The five CpG sites in a 24-base sequence were selected as the target DNA on the CDH4 gene, which is associated with colorectal cancer. When methylated and unmethylated samples hybridized with the DNA probe (HS-M24) immobilized on the electrode, an increased current signal was observed in the electrolyte containing FNC and correlated with the amount of target DNA. Furthermore, an increase in current (115 %) was observed when the PCR product of 105 bp was hybridized on the HS-M24-immobilized electrode, whereas a background level of current increase was observed in the case of unmethylated product. Such large discrimination ability might be due to the inter- and/or intra-complex formation of ferrocene with β-CD of FNC on the surface of the electrode.

DOI： 10.1002/elan.201900282

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がん診断電気チップの開発

佐藤しのぶ, 竹中繁織

Bio Clinica ( 北隆館 ) 2019年01月

新規フェロセン化ナフタレンジイミドを用いる均一溶液中でのシグナルオン型電気化学的DNase I検出査読有り

佐藤しのぶ, 福瀧修司, 竹中繁織

分析化学 ( 公益社団法人日本分析化学会 ) 68 ( 12 ) 953 - 960 2019年01月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）

A simple DNase I detecting method was achieved under a homogenous medium containing DNA duplex and a newly synthesized ferrocenylnaphthalene diimide derivative (FND) carrying alanine (1) or alanine-lysine residue (2) as an electrochemical indicator. The binding affinity of 1 or 2 with double stranded DNA was 105 M−1 or 106 M−1 order, which was estimated from a Scatchard analysis calculated by its absorption change upon the addition of calf thymus DNA (CT-DNA). It was expected that 1 or 2 intercalated to double stranded DNA with a threading mode and electrostatic interaction of the linker ammonium cations of 1 or 2 with phosphate anions of the DNA duplex backbone. The largest current increase after a DNase I treatment was observed under the mixture in a ratio of [3]:[CT-DNA-bp] = 10 : 1 (bp: DNA concentration per base pair), where the distance between the bound ligands is expected to a 10 bp or 34 Å theoretically. When considering to be ca. 30 Å of DNase I size and decreasing of the current increase under an increased amount of 3, DNase I required over ten base pairs to access and digest with the DNA duplex. The current increase in the case of a mixture of 10 μM 1 or 2 and 100 μM CT-DNA was obtained 340 % or 640 %, respectively, after being treated with DNase I (final concentration: 2.0 × 10−3 U μL−1), which might be derived from an apperarent molecular weight or diffusion coefficient of the ligand bound DNA duplex and the detection limit of DNase I in the case of 1 or 2 was 2.0 × 10−5 or 2.0 × 10−9 U μL−1, respectively, where the detection limit for 2 was 104-times higher than that for 1.

DOI： 10.2116/bunsekikagaku.68.953

CiNii Article

その他リンク： https://ci.nii.ac.jp/naid/130007783471

口腔がん早期発見のための剝離細胞診を応用した自己検診システムの開発:—適切な細胞採取法の検討— 査読有り

早川真奈, 土生学, 原口和也, 矢田直美, 佐藤しのぶ, 竹中繁織, 冨永和宏

日本口腔診断学会雑誌 ( 日本口腔診断学会 ) 32 ( 3 ) 191 - 196 2019年01月

記述言語：日本語掲載種別：研究論文（学術雑誌）

Previous studies conducted by our group confirmed that the electrochemical telomerase assay （ECTA） is sufficiently sensitive to detect oral cancer from the cells obtained by exfoliation from the whole oral cavity. In this study to develop a self-screening system for oral cancer using this method, we examined the optimal method for self-collection of exfoliated cells from the whole oral cavity. First, an interdental brush, a conventional silicone brush for the cervix, a newly developed silicone brush for the oral cavity, a sponge brush used in our previous studies, and a gargle were tested for exfoliation from the oral cavity in ten healthy volunteers. Cells exfoliated from the oral cavity were collected by a single examiner, to prevent inter-examiner variation. Then, exfoliated cells were collected from the same ten healthy volunteers by the same manner and stained using papanicolaou. Finally, to mimic self-examination for oral cancer, another ten healthy volunteers were instructed on how to perform a specially-made protocol for collecting exfoliated cells from their oral cavity with a sponge brush, which had been evaluated in the initial study as the most efficient method. All subjects collected a sufficient number of cells comparable to conventional methods of exfoliative cytology. In conclusion, the self-screening system for detecting oral cancer using ECTA with the cell collection protocol described herein could be promising.

DOI： 10.15214/jsodom.32.191

CiNii Article

CiNii Research

その他リンク： https://ci.nii.ac.jp/naid/130007742331

超分子相互作用を利用したDNAバンドリング

佐藤しのぶ, 開健亮, 竹中繁織

ナノ学会会報 ( ナノ学会 ) 2018年11月

がん診断電気チップの開発

佐藤しのぶ, 竹中繁織

Medical Science Digest ( ニュー・サイエンス社 ) 2018年10月

Membrane-based microwave-mediated electrochemical immunoassay for the in vitro, highly sensitive detection of osteoporosis-related biomarkers 査読有り国際誌

Kim H., Sato S., Takenaka S., Lee M.

Sensors (Switzerland) 18 ( 9 ) 2018年09月

記述言語：英語掲載種別：研究論文（学術雑誌）

© 2018 by the authors. Licensee MDPI, Basel, Switzerland. Highly sensitive and multiplexed in vitro detection of osteoporosis-related biochemical markers were carried out based on the membrane-based microwave-mediated electrochemical immunoassay (MMeEIA), where we can dramatically reduce the sample preparation time by shortening the incubation time of conjugation to obtain sensitive detection based on three dimensional conjugation of antibodies with target antigens in nylon membrane disk. C-terminal cross-linked telopeptide of type I collagen (CTx), Osteocalcin (OC), parathyroid hormone (PTH), and N-terminal propeptide of type I collagen (P1NP), which can be utilized to monitor the progress of osteoporosis, were quantified using their corresponding antibody immobilized in membranes. Coefficient of variations in this intra-and inter-assays were within 8.0% for all markers. When compared with data obtained from clinically used standard equipment (Roche modular E170), their coefficients of determination, R2 values, are mostly more than 0.9. They show that the results obtained from MMeEIA are in good agreement with that from the conventional clinical instruments.

DOI： 10.3390/s18092933

Scopus

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Synthesis of peptide-human telomere DNA conjugate as fluorometric imaging reagent for biological sodium ion 招待有り査読有り

SATO Shinobu, IMAICHI Yuuki, YOSHIURA Yukiko, NAKAZAWA Koji, TAKENAKA Shigeori

Analytical Sciences 2018年01月

Cyclic perylene diimide: Selective ligand for tetraplex DNA binding over double stranded DNA 査読有り国際誌

Vasimalla S., Sato S., Takenaka F., Kurose Y., Takenaka S.

Bioorganic and Medicinal Chemistry 25 ( 24 ) 6404 - 6411 2017年12月

記述言語：英語掲載種別：研究論文（学術雑誌）

© 2017 Elsevier Ltd Synthesized cyclic perylene diimide, cPDI, showed the binding constant of 6.3 × 10 6 M −1 with binding number of n = 2 with TA-core as a tetraplex DNA in 50 mM Tris-HCl buffer (pH = 7.4) containing 100 mM KCl using Schatchard analysis and showed a higher preference for tetraplex DNA than for double stranded DNA with over 10 3 times. CD spectra showed that TA-core induced its antiparallel conformation upon addition of cPDI in the absence or presence of K + or Na + ions. The cPDI inhibits the telomerase activity with IC 50 of 0.3 µM using TRAP assay which is potential anti-cancer drug with low side effect.

DOI： 10.1016/j.bmc.2017.10.014

Scopus

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The methylation status and expression of human telomerase reverse transcriptase is significantly high in oral carcinogenesis 査読有り国際誌

Haraguchi K., Yada N., Sato S., Habu M., Hayakawa M., Takahashi O., Sasaguri M., Takenaka S., Yoshioka I., Matsuo K., Tominaga K.

APMIS 125 ( 9 ) 797 - 807 2017年09月

記述言語：英語掲載種別：研究論文（学術雑誌）

© 2017 APMIS. Published by John Wiley & Sons Ltd Telomerase activity is present in most cancers and is tightly regulated by the expression of human telomerase reverse transcriptase (hTERT). Hypermethylation in the promoter region of hTERT contributes to the regulation of hTERT expression. In this study, we investigated the methylation and expression of hTERT in oral squamous cell carcinoma (OSCC), oral leukoplakia, and normal oral mucosa. Furthermore, we investigated the significance of hTERT to the clinicopathological findings of OSCC. 35 OSCC, 50 oral leukoplakia (epithelial dysplasia n = 25, squamous cell hyperplasia n = 25), and 10 normal oral mucosa samples were investigated through methylation-specific PCR. Immunohistochemistry was analyzed in 35 OSCC, 50 oral leukoplakia, and 4 normal oral mucosa samples. The methylation and expression of hTERT increased from normal oral mucosa to oral leukoplakia to OSCC. In OSCC, all samples were methylated. However, partial methylation (20%) or unmethylation (80%), but never complete methylation, was observed in normal oral mucosa. Additionally, hTERT expression correlated with cervical lymph node metastasis. These results suggested that the methylation and expression of hTERT is high in oral carcinogenesis and may play an important role in oral cancer. hTERT expression may also be predictive of cervical lymph node metastasis.

DOI： 10.1111/apm.12723

Scopus

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Oral Cancer Screening Based on Methylation Frequency Detection in hTERT Gene Using Electrochemical Hybridization Assay via a Multi-electrode Chip Coupled with Ferrocenylnaphthalene Diimide 査読有り国際誌

Haraguchi K., Sato S., Habu M., Yada N., Hayakawa M., Takahashi O., Yoshioka I., Matsuo K., Tominaga K., Takenaka S.

Electroanalysis 29 ( 6 ) 1596 - 1601 2017年03月

記述言語：英語掲載種別：研究論文（学術雑誌）

© 2017 Wiley-VCH Verlag GmbH & Co. KGaA, WeinheimFerrocenylnaphthalene diimide-based electrochemical hybridization assay via a multi-electrode chip was applied to detect the methylation frequency in the promoter region of human telomerase reverse transcriptase (hTERT) gene for clinical samples from tissues, local exfoliated oral cells from a lesion, or from entire oral cavity after their methylation specific PCRs. These methylation frequencies were increased with cancer progress as the following order: healthy volunteers, oral leukoplakia as precancerous lesion, and oral squamous cell carcinoma (OSCC). Operating characteristic analysis of the obtained current data doesn't only give excellent discrimination ability of OSCC, but also of oral leukoplakia from healthy volunteers for all samples. Sensitivity and specificity was 95 % and 90 %, respectively, which is a comparable with methods in practical use.

DOI： 10.1002/elan.201700028

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Ferrocenyl naphthalene diimides as tetraplex DNA binders 査読有り国際誌

Sato S., Takenaka S.

Journal of Inorganic Biochemistry 167 21 - 26 2017年02月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）

© 2016Seven ferrocenyl naphthalene diimide (FND) ligands were synthesized. Each had a higher affinity for tetraplex DNA than for either single- or double-stranded DNA. The FND binding affinities were > 105 M− 1 in 0.10 M AcOH-AcONa or AcOH-AcOK (pH 5.5) containing 0.10 M NaCl or KCl. The FNDs with the highest binding affinities for tetraplex DNA showed 23- or 8-times higher preference for tetraplex DNA than for single- or double-stranded DNA, respectively. The current signals generated from the seven FNDs bound to the tetraplex DNA immobilized on the electrode were found to correlate with the binding affinities of these ligands for the tetraplex DNA. Furthermore, using the telomerase repeat amplification protocol assay, the FND ligands could be categorized into three groups: (a) inhibiting both telomerase and Taq polymerase, (b) inhibiting telomerase alone, and (c) inhibiting neither telomerase nor Taq polymerase.

DOI： 10.1016/j.jinorgbio.2016.11.020

Scopus

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Cyclic ferrocenylnaphthalene diimide derivative as a new class of G-quadruplex DNA binding ligand 査読有り国際誌

Islam M., Sato S., Sato S., Shinozaki S., Takenaka S., Takenaka S.

Bioorganic and Medicinal Chemistry Letters 27 ( 2 ) 329 - 335 2017年01月

記述言語：英語掲載種別：研究論文（学術雑誌）

© 2016 Elsevier LtdTo identify an effective ligand that binds to a G-quadruplex structure but not a double-stranded DNA (dsDNA), a set of biophysical and biochemical experiments were carried out using newly synthesized cyclic ferrocenylnaphthalene diimide (cFNDI, 1) or the non-cyclic derivative (2) with various structures of G-quadruplex DNAs and dsDNA. Compound 1 bound strongly to G-quadruplexes DNAs (106 M−1 order) with diminished binding to dsDNA (104 M−1 order) in 100 mM AcOH-AcOK buffer (pH 5.5) containing 100 mM KCl. Interestingly, 1 showed an approximately 50-fold higher selectivity to mixed hybrid-type telomeric G-quadruplex DNA (K = 3.4 × 106 M−1 and a 2:1 stoichiometry) than dsDNA (K = 7.5 × 104 M−1) did. Furthermore, 1 showed higher thermal stability to G-quadruplex DNAs than it did to dsDNA with a preference for c-kit and c-myc G-quadruplex DNAs over telomeric and thrombin binding aptamers. Additionally, 1 exhibited telomerase inhibitory activity with a half-maximal inhibitory concentration (IC50) of 0.4 μM. Compound 2 showed a preference for G-quadruplex; however, the binding affinity magnitude and preference were improved in 1 because the former had a cyclic structure.

DOI： 10.1016/j.bmcl.2016.11.037

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Electrochemical hybridization assay for methylation detection of the hTERT gene connected with oral cancer screening 査読有り

Sato S., Haraguchi K., Hayakawa M., Tominaga K., Takenaka S.

Bunseki Kagaku ( 公益社団法人日本分析化学会 ) 66 ( 6 ) 437 - 443 2017年01月

担当区分：筆頭著者記述言語：日本語掲載種別：研究論文（学術雑誌）

© 2017 The Japan Society for Analytical Chemistry. The human telomerase reverse transcriptase (hTERT) gene is encoded as a catalytic unit of telomerase. Since the methylation pattern of the CpG islands in the promoter region of hTERT gene is related with cancer development, this should serve as a suitable cancer marker. Electrochemical hybridization assay was applied to this detection using ferrocenylnaphthalene diimide (FND) as a hybrid indicator. Ten different 24-meric DNA probes were designed and immobilized on the individual electrodes of multi-electrode chip, which can hybridize with sample DNAs carrying different methylation patterns: M0, M1, M1', M2, M2', M3, M3', M4, M4', or M5, where number refers to the number of methylated cytosine of CpG islands in the promoter region of hTERT gene. Aiming for simple diagnosis of oral cancer, the total exfoliated oral cells as clinical samples were obtained from oral cancer patients (oral squamous cell carcinoma, OSCC), oral leukoplakia as precancerous lesion, and healthy volunteers. The clinical samples were amplified by a methylation-specific polymerase chain reaction (MSP) with bisulfite treated genome DNA from the total exfoliated oral cells and hybridized with 10-probe DNA immobilized multi-electrode chip. The current increase ratio of M4, M4', M5 probe for detecting methylation was increased with cancer progress according to the following order: healthy volunteers, oral leukoplakia, and OSCC. Receiver operatorating characteristic curve of the obtained data gave the threshold values in the case of the detection of OSCC from all samples to obtain 91 % of the accuracy rate for total exfoliated oral cells using 96.6 % of the threshold values.

DOI： 10.2116/bunsekikagaku.66.437

Scopus

CiNii Article

CiNii Research

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Water-soluble porphyrinoids as G-quadruplex binders and telomerase inhibitors 査読有り国際誌

Ikawa Y., Katsumata S., Sakashita R., Sato S., Takenaka S., Furuta H., Furuta H.

Journal of Porphyrins and Phthalocyanines 20 ( 8-11 ) 1041 - 1048 2016年08月

記述言語：英語掲載種別：研究論文（学術雑誌）

© 2016 World Scientific Publishing Company.Water-soluble derivatives of three kinds of expanded porphyrins (N-fused pentaphyrin, hexaphyrin, and heptaphyrin) were synthesized and their binding ability to G-quadruplex (G4-) DNA and the inhibitory effects on enzymatic telomere extension were investigated. While expanded porphyrins increased the melting temperature of G4-DNA more effectively than the regular porphyrins, a porphyrin isomer (N-confused porphyrin) showed the highest inhibitory effect on telomerase activity.

DOI： 10.1142/S108842461650053X

Scopus

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DNA methylation detection based on difference of base content 招待有り査読有り

Sato S., Sato S., Ohtsuka K., Ohtsuka K., Honda S., Sato Y., Takenaka S., Takenaka S.

Journal of Physics: Conference Series 704 ( 1 ) 2016年04月

記述言語：英語掲載種別：研究論文（国際会議プロシーディングス）

Methylation frequently occurs in cytosines of CpG sites to regulate gene expression. The identification of aberrant methylation of certain genes is important for cancer marker analysis. The aim of this study was to determine the methylation frequency in DNA samples of unknown length and/or concentration. Unmethylated cytosine is known to be converted to thymine following bisulfite treatment and subsequent PCR. For this reason, the AT content in DNA increases with an increasing number of methylation sites. In this study, the fluorescein-carrying bis-acridinyl peptide (FKA) molecule was used for the detection of methylation frequency. FKA contains fluorescein and two acridine moieties, which together allow for the determination of the AT content of double-stranded DNA fragments. Methylated and unmethylated human genomes were subjected to bisulfide treatment and subsequent PCR using primers specific for the CFTR, CDH4, DBC1, and NPY genes. The AT content in the resulting PCR products was estimated by FKA, and AT content estimations were found to be in good agreement with those determined by DNA sequencing. This newly developed method may be useful for determining methylation frequencies of many PCR products by measuring the fluorescence in samples excited at two different wavelengths.

DOI： 10.1088/1742-6596/704/1/012015

Scopus

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Electrochemical approaches to the study of DNA/drug interactions 招待有り査読有り

Sato S., Takenaka S.

Methods for Studying Nucleic Acid/Drug Interactions 241 - 259 2016年04月

電気化学的手法によるDNA/薬剤相互作用解析

佐藤しのぶ、竹中繁織

DOJIN News ( DOJINDO ) 156 1 - 4 2016年03月

電気化学的口腔癌診断法の開発：フェロセン化ナフタレンジイミド誘導体を利用した電気化学的テロメラーゼ検出

佐藤しのぶ、竹中繁織

化学とマイクロ・ナノシステム学会誌 ( 一般社団法人　化学とマイクロ・ナノシステム学会 ) 15 ( 1 ) 7 - 12 2016年03月

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