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Article ; Online: Th1/17 polarization and potential treatment by an anti-interferon-γ DNA aptamer in Hunner-type interstitial cystitis.

Akiyama, Yoshiyuki / Harada, Kaori / Miyakawa, Jimpei / Kreder, Karl J / O'Donnell, Michael A / Daichi, Maeda / Katoh, Hiroto / Hori, Miyuki / Owari, Kensuke / Futami, Kazunobu / Ishikawa, Shumpei / Ushiku, Tetsuo / Kume, Haruki / Homma, Yukio / Luo, Yi

iScience

2023 Volume 26, Issue 11, Page(s) 108262

Abstract: Hunner-type interstitial cystitis (HIC) is a rare, enigmatic inflammatory disease of the urinary bladder with no curative treatments. In this study, we aimed to characterize the unique cellular and immunological factors specifically involved in HIC by ... ...

Abstract	Hunner-type interstitial cystitis (HIC) is a rare, enigmatic inflammatory disease of the urinary bladder with no curative treatments. In this study, we aimed to characterize the unique cellular and immunological factors specifically involved in HIC by comparing with cystitis induced by
Language	English
Publishing date	2023-10-21
Publishing country	United States
Document type	Journal Article
ISSN	2589-0042
ISSN (online)	2589-0042
DOI	10.1016/j.isci.2023.108262
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Article: RECQL1 and WRN DNA repair helicases: potential therapeutic targets and proliferative markers against cancers.

Futami, Kazunobu / Furuichi, Yasuhiro

Frontiers in genetics

2015 Volume 5, Page(s) 441

Abstract: RECQL1 and WRN helicases in the human RecQ helicase family participate in maintaining genome stability, DNA repair, replication, and recombination pathways in the cell cycle. They are expressed highly in rapidly proliferating cells and tumor cells, ... ...

Abstract	RECQL1 and WRN helicases in the human RecQ helicase family participate in maintaining genome stability, DNA repair, replication, and recombination pathways in the cell cycle. They are expressed highly in rapidly proliferating cells and tumor cells, suggesting that they have important roles in the replication of a genome. Although mice deficient in these helicases are indistinguishable from wild-type mice, their embryonic fibroblasts are sensitive to DNA damage. In tumor cells, silencing the expression of RECQL1 or WRN helicase by RNA interference induces mitotic catastrophe that eventually kills tumor cells at the mitosis stage of the cell cycle. By contrast, the same gene silencing by cognate small RNA (siRNA) never kills normal cells, although cell growth is slightly delayed. These findings indicate that RECQL1 and WRN helicases are ideal molecular targets for cancer therapy. The molecular mechanisms underlying these events has been studied extensively, which may help development of anticancer drugs free from adverse effects by targeting DNA repair helicases RECQL1 and WRN. As expected, the anticancer activity of conventional genotoxic drugs is significantly augmented by combined treatment with RECQL1- or WRN-siRNAs that prevents DNA repair in cancer cells. In this review, we focus on studies that clarified the mechanisms that lead to the specific killing of cancer cells and introduce efforts to develop anticancer RecQ-siRNA drugs free from adverse effects.
Language	English
Publishing date	2015-01-09
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2606823-0
ISSN	1664-8021
ISSN	1664-8021
DOI	10.3389/fgene.2014.00441
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Article: Genetic Alphabet Expansion Provides Versatile Specificities and Activities of Unnatural-Base DNA Aptamers Targeting Cancer Cells.

Futami, Kazunobu / Kimoto, Michiko / Lim, Yun Wei Shermane / Hirao, Ichiro

Molecular therapy. Nucleic acids

2018 Volume 14, Page(s) 158–170

Abstract: The potential of genetic alphabet expansion technologies using artificial extra base pairs (unnatural base pairs) has been rapidly expanding and increasing. We present that the hydrophobic unnatural base, 7-(2-thienyl)imidazo[4,5-b]pyridine (Ds), which ... ...

Abstract	The potential of genetic alphabet expansion technologies using artificial extra base pairs (unnatural base pairs) has been rapidly expanding and increasing. We present that the hydrophobic unnatural base, 7-(2-thienyl)imidazo[4,5-b]pyridine (Ds), which acts as a fifth letter in a DNA library, provides a series of high-affinity DNA aptamers with versatile binding specificities and activities to cancer cells. These Ds-containing DNA aptamers were generated by a method called cell-ExSELEX to target three breast cancer cell lines: MCF7, MDA-MB-231, and T-47D. Aptamer 14A-MCF7, which targets MCF7 cells, specifically binds to MCF7 cells, but not other cancer cell lines. Aptamer 07-MB231, which targets MDA-MB-231 cells, binds to a series of metastatic bone and lung cancer cell lines. Aptamer 05-MB231 targets MDA-MB-231 cells, but it also binds to all of the cancer and leukemia cell lines that we examined. None of these aptamers bind to normal cell lines, such as MCF10A and HUVEC. In addition, aptamers 14A-MCF7 and 05-MB231 are internalized within the cancer cells, and aptamer 05-MB231 possesses anti-proliferative properties against most cancer cell lines that we examined. These aptamers and the generation method are broadly applicable to cancer cell imaging, biomarker discovery, cancer cell profiling, anti-cancer therapies, and drug delivery systems.
Language	English
Publishing date	2018-11-29
Publishing country	United States
Document type	Journal Article
ZDB-ID	2662631-7
ISSN	2162-2531
ISSN	2162-2531
DOI	10.1016/j.omtn.2018.11.011
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Article ; Online: Quantitation of full-size small interfering RNA by tailing with terminal deoxynucleotidyl transferase and reverse transcription-polymerase chain reaction analysis.

Futami, Kazunobu / Furuichi, Yasuhiro

Analytical biochemistry

2009 Volume 385, Issue 2, Page(s) 386–388

Abstract: Accurate estimation of small interfering RNA (siRNA) concentration in cells and blood is increasingly important for pharmacokinetic studies required to develop siRNA drugs. We report a method that detects siRNA having 3'-terminal deoxynucleotide ... ...

Abstract	Accurate estimation of small interfering RNA (siRNA) concentration in cells and blood is increasingly important for pharmacokinetic studies required to develop siRNA drugs. We report a method that detects siRNA having 3'-terminal deoxynucleotide overhangs, such as 3'-dTdT, present in most chemically synthesized siRNAs. Short overhangs were elongated to oligo-dG by incubation with terminal deoxynucleotidyl transferase and dGTP and were used as priming sites for reverse transcription of siRNA to complementary DNA (cDNA). The resultant cDNA was used as a template for quantitation by polymerase chain reaction. This method was reliable for determining the pharmacokinetics of siRNA in blood of injected mice.
MeSH term(s)	Animals ; DNA Nucleotidylexotransferase/metabolism ; DNA, Complementary ; Deoxyguanine Nucleotides ; Mice ; RNA, Small Interfering/analysis ; RNA, Small Interfering/blood ; RNA, Small Interfering/pharmacokinetics ; Research Design ; Reverse Transcriptase Polymerase Chain Reaction/methods
Chemical Substances	DNA, Complementary ; Deoxyguanine Nucleotides ; RNA, Small Interfering ; deoxyguanosine triphosphate (8C2O37Y44Q) ; DNA Nucleotidylexotransferase (EC 2.7.7.31)
Language	English
Publishing date	2009-02-15
Publishing country	United States
Document type	Evaluation Studies ; Journal Article
ZDB-ID	1110-1
ISSN	1096-0309 ; 0003-2697
ISSN (online)	1096-0309
ISSN	0003-2697
DOI	10.1016/j.ab.2008.11.037
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Article ; Online: Detection of siRNA administered to cells and animals by using a fluorescence intensity distribution analysis polarization system.

Abe, Takashi / Goda, Kazuhito / Futami, Kazunobu / Furuichi, Yasuhiro

Nucleic acids research

2009 Volume 37, Issue 7, Page(s) e56

Abstract: Small interfering RNA (siRNA) has excellent pharmacological features and is expected to be used for therapeutic drug development. To this end, however, new RNA technology needs to be established so that extremely small amounts (less than 1 pmol) of siRNA ...

Abstract	Small interfering RNA (siRNA) has excellent pharmacological features and is expected to be used for therapeutic drug development. To this end, however, new RNA technology needs to be established so that extremely small amounts (less than 1 pmol) of siRNA can be detected in organs of experimental animals and in human blood to facilitate pharmacokinetics studies. An important feature is that this new technology is not dependent on radioisotopes and can detect siRNA molecules identical to those used for drug development in preclinical tests with experimental animals or in clinical tests with humans. We report a convenient method that can detect small amounts of siRNA. The method uses high-power confocal microscopic analysis of fluorescence polarization in DNA probes that are bound to one of the strands of siRNA and directly quantitates the copy number of siRNA molecule after extraction from specimens. A pharmacokinetic study to examine the blood retention time of siRNA/cationic liposomes in mice showed that this straightforward method is consistent with the other reverse transcriptase polymerase chain reaction amplification-based method. We believe that the entire process is simple and applicable for a high-throughput analysis, which provides excellent technical support for fundamental research on RNA interference and development of siRNA drugs.
MeSH term(s)	Animals ; Base Sequence ; Cell Line, Tumor ; DNA Probes/chemistry ; Fluorescence Polarization/methods ; Humans ; Injections, Intravenous ; Mice ; Microscopy, Confocal ; RNA, Small Interfering/administration & dosage ; RNA, Small Interfering/analysis ; RNA, Small Interfering/blood ; Tissue Distribution ; Transfection
Chemical Substances	DNA Probes ; RNA, Small Interfering
Language	English
Publishing date	2009-03-12
Publishing country	England
Document type	Journal Article
ZDB-ID	186809-3
ISSN	1362-4962 ; 1362-4954 ; 0301-5610 ; 0305-1048
ISSN (online)	1362-4962 ; 1362-4954
ISSN	0301-5610 ; 0305-1048
DOI	10.1093/nar/gkp131
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Zs.A 1067: Show issues

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Article ; Online: RecQL1 DNA repair helicase: A potential tumor marker and therapeutic target against hepatocellular carcinoma.

Futami, Kazunobu / Ogasawara, Sachiko / Goto, Hideyuki / Yano, Hirohisa / Furuichi, Yasuhiro

International journal of molecular medicine

2010 Volume 25, Issue 4, Page(s) 537–545

Abstract: RecQL1 in the human RecQ DNA helicase family participates in DNA repair and recombination pathways in cell cycle replication. Immunohistochemical analysis of human hepatocellular carcinoma (HCC) tissues showed that RecQL1 expression is strongly ... ...

Abstract	RecQL1 in the human RecQ DNA helicase family participates in DNA repair and recombination pathways in cell cycle replication. Immunohistochemical analysis of human hepatocellular carcinoma (HCC) tissues showed that RecQL1 expression is strongly correlated with histological grade and MIB-1 indices of HCC, and that the expression was greater in simple HCCs inducing extranodular growth or portal vein invasion than in HCCs not inducing extranodular growth or portal vein invasion. These histological data reveal the potential of RecQL1 as a biological marker predicting the malignancy and progression of liver cancer. High expression profiles were also produced by various HCC cells, including HCC cell lines established by us. When RecQL1 expression was silenced by siRNA in vitro, most HCC cells died of mitotic catastrophe. In a mouse orthotopic xenograft model of liver cancer with transplanted human HCC, RecQL1-siRNA mixed with cationic liposomes exhibited a strong anticancer effect that prevented the growth of the cancer. RecQL1-siRNA inhibited the growth of human HCC in the mouse liver, confirming that RecQL1 is an excellent molecular agent against liver cancer and suggests that RecQL1-siRNA formulated with liver-prone liposomes has excellent potential as a therapeutic drug against liver cancers.
MeSH term(s)	Animals ; Biomarkers, Tumor/metabolism ; Carcinoma, Hepatocellular/enzymology ; Carcinoma, Hepatocellular/genetics ; Carcinoma, Hepatocellular/pathology ; Carcinoma, Hepatocellular/therapy ; Cell Cycle Proteins/genetics ; Cell Cycle Proteins/metabolism ; Cell Line, Tumor ; Cell Proliferation ; DNA Repair ; Gene Expression Regulation, Neoplastic ; Gene Silencing ; Humans ; Immunohistochemistry ; Ki-67 Antigen/metabolism ; Liposomes ; Liver Neoplasms/enzymology ; Liver Neoplasms/genetics ; Liver Neoplasms/pathology ; Liver Neoplasms/therapy ; Mice ; Mitosis ; RNA, Small Interfering/administration & dosage ; RNA, Small Interfering/metabolism ; RecQ Helicases/metabolism ; Xenograft Model Antitumor Assays
Chemical Substances	Biomarkers, Tumor ; Cell Cycle Proteins ; Ki-67 Antigen ; Liposomes ; RNA, Small Interfering ; RecQ Helicases (EC 3.6.4.12)
Language	English
Publishing date	2010-03-01
Publishing country	Greece
Document type	Journal Article
ZDB-ID	1444428-8
ISSN	1791-244X ; 1107-3756
ISSN (online)	1791-244X
ISSN	1107-3756
DOI	10.3892/ijmm_00000375
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Zs.A 4942: Show issues

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Article: Mitochondrial and nuclear localization of human Pif1 helicase.

Futami, Kazunobu / Shimamoto, Akira / Furuichi, Yasuhiro

Biological & pharmaceutical bulletin

2007 Volume 30, Issue 9, Page(s) 1685–1692

Abstract: Yeast Pif1 DNA helicase is the prototype member of a helicase subfamily participating in the maintenance of telomere, ribosome, and mitochondria DNAs. The Pif1 DNA helicase family is highly conserved from yeast to human, but the biochemical nature of ... ...

Abstract	Yeast Pif1 DNA helicase is the prototype member of a helicase subfamily participating in the maintenance of telomere, ribosome, and mitochondria DNAs. The Pif1 DNA helicase family is highly conserved from yeast to human, but the biochemical nature of human homologues remains to be clarified. To this end, we investigated the transcriptional unit of human Pif1 gene and its encoded protein hPif1. The results showed that the hPif1 gene product has at least two isoforms consisting of the conserved helicase motif and differential C-terminal regions derived from alternative splicing of the gene transcript. Deletion mutant analysis showed that Pif1 helicase has nuclear localization signal and mitochondria targeting signal at the N-terminal and C-terminal regions, respectively. In HeLa cells, hPif1 helicase expression was induced by the release of cells from serum starvation, suggesting that hPif1 has roles in the S phase. Consistently, the down regulation of the hPif1 helicase by RNA interference with siRNA caused a cell cycle delay at the S phase. These findings suggest that hPif1 in the nucleus may be involved in chromosome maintenance in association with DNA replication, while the function of hPif1 remains to be clarified.
MeSH term(s)	Adenosine Triphosphatases/metabolism ; Blotting, Western ; Cell Cycle/drug effects ; Cell Nucleus/enzymology ; DNA/biosynthesis ; DNA/genetics ; DNA Helicases/genetics ; DNA Helicases/isolation & purification ; DNA Helicases/metabolism ; Flow Cytometry ; HeLa Cells ; Humans ; Microscopy, Confocal ; Mitochondria/enzymology ; RNA, Small Interfering/pharmacology ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/isolation & purification ; S Phase/drug effects ; Subcellular Fractions/drug effects ; Subcellular Fractions/metabolism
Chemical Substances	RNA, Small Interfering ; Recombinant Proteins ; DNA (9007-49-2) ; Adenosine Triphosphatases (EC 3.6.1.-) ; DNA Helicases (EC 3.6.4.-) ; PIF1 protein, human (EC 5.99.-)
Language	English
Publishing date	2007-08-29
Publishing country	Japan
Document type	Journal Article
ZDB-ID	1150271-x
ISSN	1347-5215 ; 0918-6158
ISSN (online)	1347-5215
ISSN	0918-6158
DOI	10.1248/bpb.30.1685
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Zs.A 1474: Show issues

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Article: Modulation of the expression of bloom helicase by estrogenic agents.

Iso, Takako / Futami, Kazunobu / Iwamoto, Teruaki / Furuichi, Yasuhiro

Biological & pharmaceutical bulletin

2006 Volume 30, Issue 2, Page(s) 266–271

Abstract: We report that the expression of Bloom helicase (BLM) was up-regulated by 17beta-estradiol (E2) in estrogen receptor (ER)-positive mammary tumor MCF-7 cells, but was hardly modulated in ER-negative mammary tumor MDA-MB-231 cells. ER antagonist ICI182780 ... ...

Abstract	We report that the expression of Bloom helicase (BLM) was up-regulated by 17beta-estradiol (E2) in estrogen receptor (ER)-positive mammary tumor MCF-7 cells, but was hardly modulated in ER-negative mammary tumor MDA-MB-231 cells. ER antagonist ICI182780 blocked the E2 effect on BLM expression in MCF-7 cells. From these results we conclude that ER participates in up-regulation of BLM expression in MCF-7 cells by means of E2. Similar results were obtained when MCF-7 cells were treated with bisphenol A (BPA), an endocrine-disrupting chemical having a weak estrogenic activity. The ER binding ability of BPA is estimated at 1/1000 of E2 ability, and in this study about 1000-times more BPA was needed for the same levels of estrogenic effect of E2. The expression of cell-cycle associated genes, cdc6, MCM5, MCM2, Myt1, PCNA and AuroraA were up-regulated by E2 and BPA treatment in MCF-7 cells accompanied by up-regulation of BLM. In this BLM promoter study, Sp1 elements in the upper region of BLM modulated transcription, but were not indispensable for E2 response. Our results suggested that up-regulation of BLM expression by E2 and BPA is ER-dependent and may be responsible for repair of DNA damage caused by the genotoxicity of these estrogenic agents.
MeSH term(s)	Adenosine Triphosphatases/biosynthesis ; Adenosine Triphosphatases/genetics ; Benzhydryl Compounds ; Cell Line, Tumor ; DNA Helicases/biosynthesis ; DNA Helicases/genetics ; Estradiol/pharmacology ; Estrogens/pharmacology ; Humans ; Luciferases/metabolism ; Phenols/pharmacology ; Promoter Regions, Genetic ; RNA, Messenger/biosynthesis ; RecQ Helicases ; Receptors, Estrogen/metabolism ; Up-Regulation
Chemical Substances	Benzhydryl Compounds ; Estrogens ; Phenols ; RNA, Messenger ; Receptors, Estrogen ; Estradiol (4TI98Z838E) ; Luciferases (EC 1.13.12.-) ; Adenosine Triphosphatases (EC 3.6.1.-) ; Bloom syndrome protein (EC 3.6.1.-) ; DNA Helicases (EC 3.6.4.-) ; RecQ Helicases (EC 3.6.4.12) ; bisphenol A (MLT3645I99)
Language	English
Publishing date	2006-06-14
Publishing country	Japan
Document type	Journal Article
ZDB-ID	1150271-x
ISSN	1347-5215 ; 0918-6158
ISSN (online)	1347-5215
ISSN	0918-6158
DOI	10.1248/bpb.30.266
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Article: Detection of siRNA administered to cells and animals by using a fluorescence intensity distribution analysis polarization system

Abe, Takashi / Goda, Kazuhito / Futami, Kazunobu / Furuichi, Yasuhiro

Nucleic acids research. 2009 Apr., v. 37, no. 7

2009

Abstract	Small interfering RNA (siRNA) has excellent pharmacological features and is expected to be used for therapeutic drug development. To this end, however, new RNA technology needs to be established so that extremely small amounts (less than 1 pmol) of siRNA can be detected in organs of experimental animals and in human blood to facilitate pharmacokinetics studies. An important feature is that this new technology is not dependent on radioisotopes and can detect siRNA molecules identical to those used for drug development in preclinical tests with experimental animals or in clinical tests with humans. We report a convenient method that can detect small amounts of siRNA. The method uses high-power confocal microscopic analysis of fluorescence polarization in DNA probes that are bound to one of the strands of siRNA and directly quantitates the copy number of siRNA molecule after extraction from specimens. A pharmacokinetic study to examine the blood retention time of siRNA/cationic liposomes in mice showed that this straightforward method is consistent with the other reverse transcriptase polymerase chain reaction amplification-based method. We believe that the entire process is simple and applicable for a high-throughput analysis, which provides excellent technical support for fundamental research on RNA interference and development of siRNA drugs.
Keywords	DNA probes ; RNA interference ; blood ; drugs ; fluorescence ; humans ; laboratory animals ; mice ; pharmacokinetics ; radionuclides ; reverse transcriptase polymerase chain reaction ; small interfering RNA ; technology
Language	English
Dates of publication	2009-04
Size	p. e56.
Document type	Article
ZDB-ID	186809-3
ISSN	0301-5610 ; 0305-1048
ISSN	0301-5610 ; 0305-1048
DOI	10.1093/nar/gkp131
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Role of Werner syndrome gene product helicase in carcinogenesis and in resistance to genotoxins by cancer cells.

Futami, Kazunobu / Ishikawa, Yuichi / Goto, Makoto / Furuichi, Yasuhiro / Sugimoto, Masanobu

Cancer science

2008 Volume 99, Issue 5, Page(s) 843–848

Abstract: Werner syndrome (WS) is an autosomal recessive genetic disorder causing premature aging, and WRN has been identified as the causative gene of WS. The product of the WRN gene (WRN) acts as a DNA helicase with exonuclease activity, and data have ... ...

Abstract	Werner syndrome (WS) is an autosomal recessive genetic disorder causing premature aging, and WRN has been identified as the causative gene of WS. The product of the WRN gene (WRN) acts as a DNA helicase with exonuclease activity, and data have accumulated showing that the WRN gene strongly participates in carcinogenesis: (1) the normal WRN gene likely participates in the immortalization of B-lymphoblastoid cell lines through telomeric crisis caused by telomere shortening, (2) a much higher incidence of rare cancers occurs in WS patients than in other kinds of patients, and (3) levels of WRN expressed in virus-transformed cells and cancer cells are usually markedly up-regulated and are inversely correlated with the sensitivity of these cells against various genotoxins, including camptothecin. In this paper, we review the events that show a close correlation of the WRN gene and WRN with carcinogenesis and their underlying molecular mechanisms.
MeSH term(s)	Cell Transformation, Neoplastic ; Chromosomal Instability ; DNA Helicases/genetics ; DNA Helicases/metabolism ; DNA Repair ; Exodeoxyribonucleases/genetics ; Exodeoxyribonucleases/metabolism ; Humans ; Models, Biological ; Mutagens/toxicity ; Mutation ; Neoplasms/enzymology ; Neoplasms/genetics ; RecQ Helicases/genetics ; RecQ Helicases/metabolism ; Telomere/metabolism ; Werner Syndrome/genetics ; Werner Syndrome/metabolism ; Werner Syndrome Helicase
Chemical Substances	Mutagens ; Exodeoxyribonucleases (EC 3.1.-) ; DNA Helicases (EC 3.6.4.-) ; RecQ Helicases (EC 3.6.4.12) ; WRN protein, human (EC 3.6.4.12) ; Werner Syndrome Helicase (EC 3.6.4.12)
Language	English
Publishing date	2008-05
Publishing country	England
Document type	Journal Article ; Review
ISSN	1349-7006
ISSN (online)	1349-7006
DOI	10.1111/j.1349-7006.2008.00778.x
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