Identification of Reduced Fibronectin 1 Gene Expression in Osteoblasts following Hydrogen Peroxide Treatment using Subtractive Gene Cloning
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概要
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Bone formation steadily declines with age resulting in a significant loss of bone mass, while reactive oxygen species (ROS) are thought to be major contributors to the aging process. In the present study, we attempted to identify which gene had an altered transcription level when osteoblasts were treated with ROS, H_2O_2, using a subtraction gene cloning technique. A gene clone was obtained from cDNA of cells from the osteoblastic cell line MC3T3-E1 after subtraction of the cDNA of H_2O_2-treated cells and designated as MC-ROS-7. To confirm the gene product, the DNA sequence of MC-ROS-7 was determined and homology was assessed using a nucleotide-nucleotide BLAST (BLASTN) search of the NCBI database, which revealed a 99.7% homology with fibronectin 1 (FN) gene. A significant lower level of FN mRNA was found in H_2O_2-treated cells when compared to that in non-treated cells using endpoint RT-PCR and real-time PCR assays. Our results suggest that a reduction of FN gene expression by ROS is involved in the decline of bone formation in osteoblasts during the ageing process.
著者
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ABIKO Yoshimitsu
Department of Biochemistry and Molecular Biology, Nihon University School of Dentistry at Matsudo
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Sato Toshinori
Department Of Biosciences And Informatics Faculty Of Science And Technology Keio University
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WATANABE NOBUYUKI
Department of Surgery, Kagawa Rosai Hospital
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Sato Toshinori
Department Of Artificial Intelligence Kyushu Institute Of Technology
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Abiko Yoshimitsu
Department Of Biochemistry And Molecular Biochemistry Nihon University School Of Dentistry At Matsud
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Abiko Yoshimitsu
Department Of Biochemistry And Molecular Biology Nihon University School Of Dentistry:research Insti
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Hamajima Susumu
Department of Biochemistry and Molecular Biology, Nihon University School of Dentistry
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Otsuka Yoko
Department of Dentistry for the Disabled, Nihon University School of Dentistry
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Yang Chia
Department of Biochemistry and Molecular Biology, Nihon University School of Dentistry
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Mega Junichi
Department of Dentistry for the Disabled, Nihon University School of Dentistry
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Sakae Toshiro
Research Institute of Oral Sciences, Nihon University School of Dentistry
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Sakae Toshiro
Research Institute Of Oral Sciences Nihon University School Of Dentistry:department Of Histology Cyt
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Mega Junichi
Department Of Dentistry For The Disabled And Research Institute Of Oral Science Nihon University Sch
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Mega Junichi
Department Of Dentistry For The Disabled Nihon University School Of Dentistry:research Institute Of
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Yang Chia
Department Of Biochemistry And Molecular Biology Nihon University School Of Dentistry
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Hamajima Susumu
Department Of Biochemistry And Molecular Biology Nihon University School Of Dentistry
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Otsuka Yoko
Department Of Dentistry For The Disabled Nihon University School Of Dentistry:research Institute Of
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Sato Toshinori
Department Of Dentistry For The Disabled Nihon University School Of Dentistry
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Watanabe Nobuyuki
Department Of Biochemistry And Molecular Biology Nihon University School Of Dentistry
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Sato Toshinori
Department of Applied Chemistry, Keio University
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Otsuka Tanaka
Department of Dentistry for the Disabled, Nihon University School of Dentistry
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