肝細胞の分裂寿命からみた肝不全機構の解明と治療戦略への応用

• 研究課題/領域番号: 14570156
• 研究種目: 基盤研究(C)
• 配分区分: 補助金
• 応募区分: 一般
• 研究分野: 人体病理学
• 研究機関: 岩手医科大学
• 研究代表者: 増田 友之 岩手医科大学, 医学部, 教授 (10199698)
• 研究分担者: 及川 浩樹 岩手医科大学, 医学部, 助手 (50285582) ; 前沢 千早 岩手医科大学, 医学部, 講師 (10326647)
• 研究期間 (年度): 2002 – 2003
• 研究課題ステータス: 完了 (2003年度)
• 配分額: 3,500千円 (直接経費: 3,500千円); 2003年度: 1,200千円 (直接経費: 1,200千円); 2002年度: 2,300千円 (直接経費: 2,300千円)
• キーワード: 幹細胞 / hTERT / TRE assay / 分裂寿命 / エストロゲン / 生体内分子間相互作用 / テロメア / テロメラーゼ / 肝臓 / 肝不全

研究概要

当該研究課題では,テロメア長およびテロメアの伸長機構に関連するテロメラーゼ分子に注目して,(1)肝炎・肝硬変患者における肝細胞残存分裂回数の分子診断法の開発,ならびに,(2)テロメラーゼ誘導による肝細胞寿命の延長に関する基礎的検討を試みた.
研究期間内に以下の成果を達成した.
(1)生体内分子間相互作用解析装置(BIACORE 3000,BIACORE AB, Uppsala, Sweden)を用いてテロメラーゼ活性を定量的に解析するtelomeric repeat elongation (TRE) assayを開発した.TRE assayはPCR法によらない優れたテロメラーゼ活性の評価法であることが明らかとなった.
(2)エストロゲンのテロメラーゼ誘導による,肝細胞分裂寿命の延長に関する基礎的検討をヒト肝培養細胞株(Hc-cells, hNheps, WRL)を用いて行った.テロメラーゼ活性は2-4倍に上昇した.テロメラーゼ活性の上昇は,hTRET hTERT (human telomerase reverse transcriptase)の発現増強によるものであった.さらにエストロゲンの長期添加実験では,添加群でテロメア長の短縮が有意に抑制され,senescence-associated β-galactosidase陽性細胞比率も減少していた.
(3)四塩化炭素肝硬変モデルラットを作製し,外因性エストロゲン投与によるテロメア長の短縮抑制効果を検討した.エストロゲン投与群で有意にテロメア長短縮が抑制されていた(文献22).
以上より,肝細胞の分裂寿命は外因性のエストロゲン投与により延長させることが可能であり,新たな慢性肝疾患患者の治療戦略の可能性が示唆された.

研究成果

• [文献書誌] Maesawa C, Iijima S, Sato N, Yoshinori N, Suzuki M, Tarusawa M, Ishida K, Tamura G, Saito K, Masuda T.: "Esophageal extraskeletal Ewing's sarcoma."Hum Pathol.. 33(1). 130-132 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Tarusawa M, Yashima A, Endo M, Maesawa C.: "Quantitative assessment of minimal residual disease in childhood lymphoid malignancies using an allele-specific oligonucleotide real-time quantitative polymerase chain reaction."Int J Hematol.. 75(2). 166-173 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Iijima S, Maesawa C, Sato N, Ikeda K, Inaba T, Akiyama Y, Ishida K, Saito K, Masuda T.: "Gastrointestinal stromal tumour of the oesophagus : significance of immunohistochemical and genetic analyses of the c-kit gene."Gastroenterol Hepatol.. 14(4). 445-448 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Wada K, Maesawa C, Satoh T, Akasaka T, Masuda T.: "A case of primary cutaneous CD30+ T-cell lymphoproliferative disorder with features of granulomatous slack skin disease."Br J Dermatol.. 147(5). 998-1002 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Ishikawa K, Koyama T, Masuda T.: "Prevalence of HBV genotypes in asymptomatic carrier residents and their clinical characteristics during long-term follow-up : the relevance to changes in the HBeAg/anti-HBe system."Hepatol Res.. 24(1). 1 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Satoh M, Nakamura M, Saitoh H, Satoh H, Akatsu T, Iwasaka J, Masuda T, Hiramori K.: "Aldosterone synthase (CYP11B2) expression and myocardial fibrosis in the failing human heart."Clin Sci (Lond).. 102(4). 381-386 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Nakamura M, Sotoh M, Kowada S, Satoh H, Tashiro A, Sato F, Masuda T, Hiramori K.: "Reversible restrictive cardiomyopathy due to light-chain deposition disease."Mayo Clin Proc.. 77(2). 193-196 (2002)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Maesawa C, Inaba T, Sato H, Iijima S, Ishida K, Terashima M, Sato R, Suzuki M, Yashima A, Ogasawara S, Oikawa H, Sato N, Saito K, Masuda T.: "A rapid biosensor chip assay for measuring of telomerase activity using surface plasmon resonance."Nucleic Acids Res.. 31(2). E4-E4 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Yashima A, Maesawa C, Uchiyama M, Tarusawa M, Satoh T, Satoh M, Enomoto S, Sugawara K, Numaoka H, Murai K, Utsugisawa T, Ishida Y, Masuda T.: "Quantitative assessment of contaminating tumor cells in autologous peripheral blood stem cells of B-cell non-Hodgkin lymphomas using immunoglobulin heavy chain gene allele-specific oligonucleotide real-time quantitative-polymerase chain reaction."Leuk Res.. 27(10). 925-934 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Nakatani K, Hagihara S, Sando S, Sakamoto S, Yamaguchi K, Maesawa C, Saito I.: "Induction of a remarkable conformational change in a human telomeric sequence by the binding of naphthyridine dimer : inhibition of the elongation of a telomeric repeat by telomerase."J Am Chem Soc.. 125(3). 662-666 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Itabashi T, Ishida Y, Masuda T.: "Consensus primers for detecting monoclonal immunoglobulin heavy chain rearrangement in B-cell lymphomas."J Clin Pathol.. 56(10). 778-779 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Itabashi T, Satoh T, Ishida Y, Masuda T.: "Development of immunoglobulin variable heavy-chain gene consensus probes with conjugated 3'-minor groove binder groups for monitoring minimal residual disease in childhood acute lymphoblastic leukemia."J Clin Pathol.. 56(12). 952-955 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Akiyama Y, Maesawa C, Ogasawara S, Terashima M, Masuda T.: "Cell-type-specific repression of the maspin gene is disrupted frequently by demethylation at the promoter region in gastric intestinal metaplasia and cancer cells."Am J Pathol.. 163(5). 1911-1919 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Tarusawa M, Satoh S, Ishida Y, Ito S, Murai K, Enomoto S, Utsugisawa T, Motoyoshi K, Masuda T.: "Development of consensus fluorogenically labeled probes of the immunoglobulin heavy-chain gene for detecting minimal residual disease in B-cell non-Hodgkin lymphomas."Cancer Sci.. 94(10). 877-885 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Rai T, Ohira H, Tojo J, Takiguchi J, Shishido S, Sato Y, Nozawa Y, Masuda T.: "A case of hepatic inflammatory pseudotumor with primary biliary cirrhosis."Hepatol Res.. 26(3). 249-253 (2003)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Ogasawara S, Maesawa C, Yamamoto M, Akiyama Y, Wada K, Fujisawa K, Higuchi T, Tomisawa Y, Sato N, Endo S, Saito K, Masuda T.: "Disruption of cell-type-specific methylation at the Maspin gene promoter is frequently involved in undifferentiated thyroid cancers."Oncogene.. 23(5). 1117-1124 (2004)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Wada K, Maesawa C, Akasaka T, Masuda T.: "Aberrant expression of the maspin gene associated with epigenetic modification in melanoma cells."J Invest Dermatol.. 122(3). 805-811 (2004)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Oyoma M, Terashima M, Takagane A, Maesawa C.: "Prognostic significance of peritoneal minimal residual disease in detecting mRNA in gastric cancer using quantitative real-time reverse transcriotive-polymerase chain reaction of carcinoembryonic antigen."Br J Surg.. 91(4). 435-443 (2004)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Itabashi T, Maesawa C, Uchiyama M, Higuchi T, Masuda T.: "Quantitative detection of mutant alleles of the K-ras gene with minor groove binder-conjugated fluorogenic DNA probes."Int J Oncol.. 24(3). 687-696 (2004)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Satoh H, Nakamura M, Satoh M, Nakajima T, Izumoto H, Maesawa C, Kawazoe K, Masuda T, Hiramori K.: "Expression and localization of Tumor Necrosis Factor-alpha and its converting enzyme in human abdominal aortic aneurysm."Clin Sci (Lond).. 106(3). 301-306 (2004)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Akiyama Y, Maesawa C, Wada K, Fujisawa K, Itabashi T, Noda Y, Honda T, Sato N, Ishida K, Takagane A, Saito K, Masuda T.: "Human PinX1, a potent telomerase inhibitor, is not involved in human gastrointestinal tract carcinoma."Oncol Rep.. 11(4). 871-874 (2004)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Sato R, Maesawa C, Fujisawa K, Wada K, Oikawa K, Takikawa Y, Suzuki K, Oikawa H, Ishikawa K, Masuda T.: "Prevention of critcal telomere shortening by oestradiol in human normal hepatic cultured cells and carbon tetrachloride induced rat liver fibrosis."Gut.. 53(7) (in press). 1001-1009
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Maesawa C, Iijina S, Sato N, Yoshinori N, Suzuki M, Tanisawa M, Ishida K, Tamura G, Saito K, Masuda T.: "Esophageal extraskeletal Ewing's sarcoma."Hum Pathol.. 33(1). 130-132 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Tarusawa M, Yashima A, Endo M, Maesawa C.: "Quantitative assessment of minimal residual disease in childhood lymphoid malignancies using an allele-specific oligonucleotide real-time quantitative polymerase chain reaction."Int J Hematol.. 75(2). 166-173 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Iijima S, Maesawa C, Sato N, Ikeda K, Inaba T, Akiyama Y, Ishida K, Saito K, Masuda T.: "Gastrointestinal stromal tumour of the oesophagus: significance of immunohistochemical and genetic analyses of the c-kit gene."Eur J Gastroenterol Hepatol.. 14(4). 445-448 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Wada K, Maesawa C, Satoh T, Akasaka T, Masuda T.: "A case of primary cutaneous CD30+ T-cell lyimphoproliferative disorder with features of granulomatous slack skin disease."Br J Dermatol.. 147(5). 998-1002 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Ishikawa K, Koyama T, Masuda T.: "Prevalence of HBV genotypes in asymptomatic carrier residents and their clinical characteristics during long-term follow-up: the relevance to changes in the HBeAg/anti-HBe system."Hepatol Res.. 24(1). 1 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Satoh M, Nakamura M, Saitoh H, Satob H, Akatsu T, Iwasaka J, Masuda T, Hiramori K.: "Aldosterone synthase (CYP11B2) expression and myocardial fibrosis in the failing human heart."Clin Sci (Lond).. 102(4). 381-386 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Nakamura M, Satoli M, Kowada S, Satoh H, Tashiro A, Sato F, Masuda T, Hiramori K.: "Reversible restrictive cardiomyopathy due to light-chain deposition disease."Mayo Clin Proc.. 77(2). 193-196 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Maesawa C, Inaba T, Sato H, Iijima S, Ishida K, Terashima M, Sato R, Suzuki M, Yashima A, Ogasawara S, Oikawa H, Sato N, Saito K, Masuda T.: "A rapid biosensor chip assay for measuring of telomerase activity using surface plasmon resonance."Nucleic Acids Res.. 31(2). E4-E4 (2002)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Yashima A, Maesawa C, Uchiyama M, Tarusawa M, Satoh T, Satoh M, Enomoto S, Sugawara K, Numaoka H, Murai K, Utsugisawa T, Ishida Y, Masuda T.: "Quantitative assessment of contaminating tumor cells in autologous peripheral blood stem cells of B-cell non-Hodgkin lymphomas using immunoglobulin heavy chain gene allele-specific oligonucleotide real-time quantitative-polymerase chain reaction."Leuk Res.. 27(10). 925-934 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Nakatani K, Hagihara S, Sando S, Sakamoto S, Yamaguchi K, Maesawa C, Saito I.: "Induction of a remarkable conformational change in a human telomeric sequence by the binding of naphthyridine dimer inhibition of the elongation of a telomeric repeat by telomerase."J Am Chem Soc.. 125(3). 662-666 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Itabashi T, Ishida Y Masuda T.: "Consensus primers for detecting monoclonal immunoglobulin heavy chain rearrangement in B-cell lymphomas."J Chin Pathol.. 56(10). 778-779 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Itabashi T, Satoh T, Ishida Y, Masuda T.: "Development of immunoglobulin variable heavy-chain gene consensus probes with conjugated 3'-minor groove binder groups for monitoring minimal residual disease in childhood acute lymphoblastic leukemia."J Chin Pathol.. 56(12). 952-955 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Akiyama Y, Maesawa C, Ogasawara S, Terashima M, Masuda T.: "Cell-type-specific repression of the maspin gene is disrupted frequently by demethylation at the promoter region in gastric intestinal metaplasia and cancer cells."Am J Pathol.. 163(5). 191-1919 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Tarusawa M, Satoh S, Ishida Y, Ito S, Murai K, Enomoto S, Utsugisawa T, Motoyoshi K, Masuda T.: "Development of consensus fluorogenically labeled probes of the immunoglobulin heavy-chain gene for detecting minimal residual disease in B-cell non-Hodgkin lymphomas."Cancer Sci.. 94(10). 877-885 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Rai T, Ohira H, Tojo J, Takiguchi J, Shishido S, Sato Y, Nozawa Y, Masuda T.: "A case of hepatic inflammatory pseudotumor with primary biliary cirrhosis."Hepatol Res.. 25(3). 249-253 (2003)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Ogasawara S, Maesawa C, Yamamoto M, Akiyama Y, Wada K, Fujisawa K, Higuchi T, Tomisawa Y, Sato N, Endo S, Saito K, Masuda T.: "Disruption of cell-type-specific methylation at the Maspin gene promoter is frequently involved in undifferentiated thyroid cancers."Oncogene.. 23(5). 1117-1124 (2004)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Wada K, Maesawa C, Akasaka T, Masuda T.: "Aberrant expression of the maspin gene associated with epigenetic modification in melanoma cells."J Invest Dermatol.. 112(3). 805-811 (2004)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Oyama M, Terashima M, Takagane A, Maesawa C.: "Prognostic significance of peritoneal minimal residual disease in detecting mRNA in gastric cancer using quantitative real-time reverse transcriotive-polymerase chain reaction of carcinoembryonic antigen."Br J Surg.. 91(4). 435-443 (2004)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Itabashi T, Maesawa C, Uchiyama M, Higuchi T, Masuda T.: "Quantitative detection of mutant alleles of the K-ras gene with minor groove binder-conjugated fluorogenic DNA probes."Int J Oncol.. 24(3). 687-696 (2004)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Satoh H, Nakamura M, Satoh M, Nakajima T, Izumoto H, Maesawa C, Kawazoc K, Masuda T, Hiramori K.: "Expression and localization of Tumor Necrosis Factor-alpha and its converting enzyme in human abdominal aortic aneurysm."Clin Sci (Lond).. 106(3). 301-306 (2004)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Akiyama Y, Maesawa C, Wada K, Fujisawa K, Itabashi T, Noda Y, Honda T, Sato N, Ishida K, Takagane A, Saito K, Masuda T.: "Human PinX1, a potent telomerase inhibitor, is not involved in human gastrointestinal tract carcinoma."Oncol Re. 11(4). 871-874 (2004)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Sato R, Maesawa C, Fujisawa K, Wada K, Oikawa K, Takikawa Y, Suzuki K, Oikawa H, Ishikawa K, Masuda T.: "Prevention of critcal telomere shortening by oestradiol in human normal hepatic cultured cells and carbon tetrachioride induced rat liver fibrosis."Gut July. 53(7)(in press). 1001-1009
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Sato R, Maesawa C, Fujisawa K, Wada K, Oikawa K, Takikawa Y, Suzuki K, Oikawa H, Ishikawa K, Masuda T: "Prevention of critical telomere shortening by estradiol in human normal hepatic cultured cells and CCl4-induced rat liver fibrosis."Gut. 53(in press). (2004)
• [文献書誌] Uchiyama M, Maesawa C.Yashima A, Tarusawa M, Satoh T, Ishida Y, Ito S, Murai K, Enomoto S, Utsugisawa T, Masuda T.: "Short consensus probes with 3-minor groove binder of the immunoglobulin heavy-chain gene for real-time quantitative PCR in B-cell non-Hodgkin lymphomas."Lab Invest. 84(in press). (2004)
• [文献書誌] Wada K, Maesawa C, Akasaka T, Masuda T: "Aberrant expression of the maspin gene associated with epigenetic modification in melanoma cells."J Invest Dermatol. 122(in press). (2004)
• [文献書誌] Ogasawara S, Maesawa C, Yamamofo M, Akiyama Y, Wada K, Fujisawa K, Higuchi T, Tomisawa Y, Sato N, Endo S, Saito K, Masuda T: "Disruption of cell-type-specific methylation at the Maspin gene promoter is frequently involved in undifferentiated thyroid cancers."Oncogene. 23. 1117-1124 (2004)
• [文献書誌] Akiyama Y, Maesawa C, Ogasawara S, Terashima M, Masuda T: "Cell-type-specific repression of the maspin gene is disrupted frequently by demethylation at the promoter region in gastric intestinal metaplasia and cancer cells."Am J Pathol. 163. 1911-1919 (2003)
• [文献書誌] Uchiyama M, Maesawa C, Yashima A, Tarusawa M, Satoh T, Ishida Y, Ito S, Murai K, Enomoto S, Utsugisawa T, Motoyoshi K, Masuda T: "Development of consensus fluorogenically labeled probes of the immunoglobulin heavy-chain gene for detecting minimal residual disease in B-cell non-Hodgkin lymphomas."Cancer Sci. 94. 875-885 (2003)
• [文献書誌] Wada K, Maesawa C, Satoh T, Akasaka T, Masuda T.: "A case of primary cutaneous CD30+ T-cell lymphoproliferative disorder with features of granulomatous slack skin disease"Br J Dermatol. 147. 998-1002 (2002)
• [文献書誌] Tarusawa M, Yoshima A, endo M, Maesawa C.: "Quantitative assessment of minimal residual disease in childhood lymphold malignancies using an allele-specific oligonucleotide real-time quantitative polymerase chain reaction"Int J Hematol. 75. 166-173 (2002)
• [文献書誌] Oikawa H, Masuda T, Kawaguchi J, Sato R.: "Three-dimensional examination of hepatic stellate cells in rat liver and response to endothelin-1 using confocal laser scanning microscopy"J Gastroenterol Hepatol. 17. 861-872 (2002)
• [文献書誌] Nakatani K, Hagihara S, Sando s, Sakamoto S, Yamaguchi K, Maesawa C, Saito I.: "Induction of a remarkable conformational change in a human telomeric sequence by the binding of naphthyridine dimer : inhibition of the elongation of a telomeric repeat by telomerase"J Am Chem Soc. 125. 662-666 (2003)
• [文献書誌] Maesawa C, Inaba T, Sato H, Iijima S, Ishida K, Trerashima M, Sato R, et al.: "A rapid biosensor chip assay for measuring of telomerase activity using surface plasmon resonance"Nucleic Acids Res. 31. E4-4 (2003)
• [文献書誌] Ishikawa K, Koyama T, Masuda T.: "Prevalence of HBV genotypes in asymptomatic carrier residents and their clinical characteristics during long-term follow-up : the relevance to changes in the HBeAg/anti-HBe system"Hepatol Res. 24. 1 (2002)