ID | 110173 |
Title Alternative | 肝細胞癌におけるDes-gamma-carboxy prothrombinの産生はpoly-(ADP-ribose) polymerase-1によりプロトロンビン遺伝子の転写が亢進することによっておこる
DCP Production Mechanism in HCC
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Author |
Kishi, Kazuhiro
Tokushima University
Tanaka, Takahiro
Tokushima University
Tomonari, Tetsu
Tokushima University
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Okamoto, Koichi
Tokushima University
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Sogabe, Masahiro
Tokushima University
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Miyamoto, Hiroshi
Tokushima University
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Okahisa, Toshiya
Tokushima University
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Muguruma, Naoki
Tokushima University
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Kajimoto, Mayumi
Tokushima University
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Keywords | des-gamma-carboxy prothrombin
PARP-1
hepatocellular carcinoma
tumor marker
Biomarkers
Prothrombin
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Content Type |
Thesis or Dissertation
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Description | Background and Aim: Although des-gamma-carboxy prothrombin (DCP) is a well-known tumor marker for hepatocellular carcinoma (HCC), the mechanism of DCP production is unclear. This study aimed to investigate the mechanism how DCP is produced in HCC cells. Methods: Levels of mRNA and DCP were analyzed by real-time polymerase chain reaction and electro-chemiluminescence immunoassay respectively. Secreted alkaline phosphatase (SEAP) expression vectors including deletion mutants of the prothrombin gene promoter were constructed for reporter gene assay. The transcription factors bound to DNA fragments were analyzed by mass spectrometry. An electrophoretic mobility shift assay (EMSA) was performed using a biotin end-labeled DNA. Results: The prothrombin mRNA levels in all 5 DCP producing cell lines were appreciably high. However, those in 2 DCP non-producing cell lines were below detectable levels. A SEAP vector with -2985 to +27 showed a very high transcription activity in DCP-producing Huh-1 cells. However, transcription abruptly decreased when the vector with -2955 to +27 was transfected, and then remained at the similar levels with larger deletion mutants, indicating the existence of a cis-element at -2985 to -2955 (31-bp). Mass spectrometry analysis identified the protein that bound to the 31-bp DNA as poly-(ADP-ribose) polymerase-1 (PARP-1). Knockdown of the PARP-1 gene by small interfering RNA in Huh-1 cells induced marked inhibition of prothrombin gene transcription. The EMSA clearly showed that PARP-1 specifically binds to the 31-bp DNA fragment in the prothrombin gene promoter. Conclusions: Our data suggest that PARP-1 activates prothrombin gene transcription and that the excessive prothrombin gene transcription induces DCP production in DCP-producing HCC cells.
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Journal Title |
Digestion
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ISSN | 00122823
14219867
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NCID | AA00628636
AA12781324
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Publisher | Karger AG
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Volume | 95
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Issue | 3
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Start Page | 242
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End Page | 251
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Published Date | 2017-04-07
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Remark | 内容要旨・審査要旨・論文本文の公開:
内容要旨・審査要旨:LID201706272001.pdf 論文本文 : k3095_fulltext.pdf 著者の申請により要約(2017-06-27公開)に替えて論文全文を公開(2018-06-15) 本論文は, 著者Tatsuya Taniguchiの学位論文として提出され, 学位審査・授与の対象となっている。 |
Rights | © 2017 S. Karger AG, Basel
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EDB ID | |
DOI (Published Version) | |
URL ( Publisher's Version ) | |
FullText File | |
language |
eng
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TextVersion |
ETD
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MEXT report number | 甲第3095号
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Diploma Number | 甲医第1339号
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Granted Date | 2017-05-25
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Degree Name |
Doctor of Medical Science
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Grantor |
Tokushima University
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departments |
Medical Sciences
University Hospital
Technical Support Department
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