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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
Author
Kishi, Kazuhiro Tokushima University
Tanaka, Hironori Tokushima University
Kajimoto, Mayumi Tokushima University
Keywords
des-gamma-carboxy prothrombin
PARP-1
hepatocellular carcinoma
tumor marker
Biomarkers
Prothrombin
Content Type
Thesis or Dissertation
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.
Journal Title
Digestion
ISSN
00122823
14219867
NCID
AA00628636
AA12781324
Publisher
Karger AG
Volume
95
Issue
3
Start Page
242
End Page
251
Published Date
2017-04-07
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
EDB ID
DOI (Published Version)
URL ( Publisher's Version )
FullText File
language
eng
TextVersion
ETD
MEXT report number
甲第3095号
Diploma Number
甲医第1339号
Granted Date
2017-05-25
Degree Name
Doctor of Medical Science
Grantor
Tokushima University
departments
Medical Sciences
University Hospital
Technical Support Department