1997 年度実績報告書

ヒト胎児性癌細胞から得られた新たな分化関連分子の機能の解明と診断・治療への応用-新たに単離された分化初期関連遺伝子EATの機能解析を中心に-

研究課題

研究課題/領域番号	08457066
研究機関	慶應義塾大学
研究代表者	秦順一慶應義塾大学, 医学部, 教授 (90051614)
研究分担者	山田健人慶應義塾大学, 医学部, 助手 (60230463) 梅澤明弘慶應義塾大学, 医学部, 講師 (70213486)
キーワード	ヒト胎児性癌細胞 / EAT遺伝子 / G3細胞 / Bcl-2 / 10(1)細胞 / モノクローナル抗体 / 胚細胞腫瘍 / CDDP
研究概要	多分化能を有するヒトEC細胞,NCR-G3細胞の分化を制御する遺伝子の単離を試み,新たな遺伝子EATを得た.本遺伝子は細胞死阻害作用を有するbcl-2ファミリーのホモロジードメインを有している.EAT遺伝子のin vitroおよび個体での機能を明らかにし,以下の結果を得た. 1)EATに対する特異抗体を作成し,ヒト組織での分布を検討した結果,胃腸管,気管支上皮,腎尿細管など単層上皮のほか,膵ラ氏島β細胞,副腎髄質などに発現がみられた.このような発現は胎生期から保存されていた. 2)EATがアポトーシスに対して抑制的に機能することを証明するため,マウス線維芽細胞10(1)のEAT安定発現株に対して各種抗癌剤を投与した結果,シスプラチンによるアポトーシスを抑制した.さらに,EAT遺伝子が導入されたトランスジェニックマウス(EAT-Tg)にシスプラチンを投与した結果,腎尿細管上皮のアポトーシスが抑制された. 3)EAT-Tgを2系統安定して保持している.Tgは10週以後インスリン産生細胞優位の著明な膵ラ氏島過形成が認められた.成人ではラ氏島はアポトーシスによってその容積を調整されており,EATの抑制作用によって過形成が惹起されたものと考えられる.飢餓状態下でグルコース投与実験を行ったところ,有意に血清インシュリン値が上昇し,この過形成が機能的であることが証明された. 4)マウスのEATホモローグ(mEAT)を独自に単離した.同分子を用いてマウス胎児性癌細胞の発現を解析した結果,ヒトEC細胞と同様の動態を示した. 以上の結果から,EATはアポトーシスを抑制することによってヒトの初期分化に寄与していることが示唆された.

研究成果
(19件)

すべてその他

すべて文献書誌 (19件)

[文献書誌] Hiraoka,N: "Establishment of three monoclonal antibodies specifc for humna developing prospermatogonia and intratubular germ cells (ITMGCs)" Lab Invest. 76. 427-438 (1997)
[文献書誌] Sugimoto,T: "Neurogenic potential of Ewing's sarcoma cells." Virchow Archiv. 430. 41-46 (1997)
[文献書誌] Ogata,T.: "Impaired male sex development in an infnat with molecularly defined partial 9p monosomy : implication for a testis forming gene (s) on 9p." J Med Genet. 34. 331-334 (1997)
[文献書誌] Kikuchi,H.: "Do intronic mutations affecting splicing of WT1 exon 9 cause Frasier syndrome?" J Med Genetics. 35. 45-48 (1998)
[文献書誌] Hiraoka,Y: "Isolation and expression of a human of human SRY-related cDNA,hSOX20." BBA. (in press). (1998)
[文献書誌] Ohkita,H: "Up-regulated expression of murine EAT,a BCL-2 related gene,in the early stage of differentiaiton of murine embryonal carcinoma cells and embryonic stem cells." BBA. (in press). (1998)
[文献書誌] Yamada,T.: "Apoptosis of human embryonal carcinoma cells with in vitro differentiation." Cell Structure Function. 21 :. 53-61, (1996)
[文献書誌] Urano,F.: "The novel chimera gene between EWS and E1A-F encoding the adenovirus E1A enhancer-binding protein in extraosseous Ewing's sarcoma" BBRC. 219 :. 608-612 (1996)
[文献書誌] Maruyma,T.: "Heat shock induces differentiation of human embryonal cacinoma cells into trophoectodermal lineages." Exp.Cell Res.224 :. 123-127 (1996)
[文献書誌] Umezawa,A.: "Involvement of mcll/EAT,Bcl-2 related gene,in differentiation of human embryonal carcinoma cells." Cell Structure Function. 21 :. 143-150, (1996)
[文献書誌] Andrews,P.W.: "A comparative analysis of cell surface antigens expressed by cell lines derived from human germ cell tumors." Int J Cancer. 66 :. 806-816, (1996)
[文献書誌] Oda,H.: "Somatic mutations of the APC gene in sporadic hepatobastokmas." Cancer Res. 56 :. 3320-3323, (1996)
[文献書誌] Hachitanda,Y.: "Stage IVS neuroblastoma : A clinical,histological and biological analysis of 45 case." Human Pathol. 27 :. 1135-38, (1996)
[文献書誌] Hiraoka,N: "Establishment of three monoclonal antibodies specifc for humna developing prospermatogonia and intratubular germ cells (ITMGCs)" Lab Invest. 76 :. 427-438, (1997)
[文献書誌] Sugimoto,T: "Neurogenic potential of Ewing's sarcoma cells." Virchow Archiv. 430 :. 41-46, (1997)
[文献書誌] Ogata,T.: "Impaired male sex development in an infnat with molecularly defined partial 9p monosomy : implication for a testis forming gene (s) on 9p." J Med Genet. 34 :. 331-334, (1997)
[文献書誌] Kikuchi,H.: "Do intronic mutations affecting splicing of WT1 exon 9 cause Frasier syndrome?" J Med Genetics. 35 :. 45-48, (1998)
[文献書誌] Hiraoka,Y.: "Isolation and expression of a human of human SRY-related cDNA,hSOX20." BBA. (in press). (1998)
[文献書誌] Ohkita,H.: "Up-regulated expression of murine EAT,a BCL-2 related gene,in the early stage of differentiaiton of murine embryonal carcinoma cells and embryonic stem cells." BBA. (in press). (1998)

1997 年度 実績報告書

ヒト胎児性癌細胞から得られた新たな分化関連分子の機能の解明と診断・治療への応用-新たに単離された分化初期関連遺伝子EATの機能解析を中心に-

研究代表者

秦 順一 慶應義塾大学, 医学部, 教授 (90051614)

研究成果

[文献書誌] Hiraoka,N: "Establishment of three monoclonal antibodies specifc for humna developing prospermatogonia and intratubular germ cells (ITMGCs)" Lab Invest. 76. 427-438 (1997)

[文献書誌] Sugimoto,T: "Neurogenic potential of Ewing's sarcoma cells." Virchow Archiv. 430. 41-46 (1997)

[文献書誌] Ogata,T.: "Impaired male sex development in an infnat with molecularly defined partial 9p monosomy : implication for a testis forming gene (s) on 9p." J Med Genet. 34. 331-334 (1997)

[文献書誌] Kikuchi,H.: "Do intronic mutations affecting splicing of WT1 exon 9 cause Frasier syndrome?" J Med Genetics. 35. 45-48 (1998)

[文献書誌] Hiraoka,Y: "Isolation and expression of a human of human SRY-related cDNA,hSOX20." BBA. (in press). (1998)

[文献書誌] Ohkita,H: "Up-regulated expression of murine EAT,a BCL-2 related gene,in the early stage of differentiaiton of murine embryonal carcinoma cells and embryonic stem cells." BBA. (in press). (1998)

[文献書誌] Yamada,T.: "Apoptosis of human embryonal carcinoma cells with in vitro differentiation." Cell Structure Function. 21 :. 53-61, (1996)

[文献書誌] Urano,F.: "The novel chimera gene between EWS and E1A-F encoding the adenovirus E1A enhancer-binding protein in extraosseous Ewing's sarcoma" BBRC. 219 :. 608-612 (1996)

[文献書誌] Maruyma,T.: "Heat shock induces differentiation of human embryonal cacinoma cells into trophoectodermal lineages." Exp.Cell Res.224 :. 123-127 (1996)

[文献書誌] Umezawa,A.: "Involvement of mcll/EAT,Bcl-2 related gene,in differentiation of human embryonal carcinoma cells." Cell Structure Function. 21 :. 143-150, (1996)

[文献書誌] Andrews,P.W.: "A comparative analysis of cell surface antigens expressed by cell lines derived from human germ cell tumors." Int J Cancer. 66 :. 806-816, (1996)

[文献書誌] Oda,H.: "Somatic mutations of the APC gene in sporadic hepatobastokmas." Cancer Res. 56 :. 3320-3323, (1996)

[文献書誌] Hachitanda,Y.: "Stage IVS neuroblastoma : A clinical,histological and biological analysis of 45 case." Human Pathol. 27 :. 1135-38, (1996)

[文献書誌] Hiraoka,N: "Establishment of three monoclonal antibodies specifc for humna developing prospermatogonia and intratubular germ cells (ITMGCs)" Lab Invest. 76 :. 427-438, (1997)

[文献書誌] Sugimoto,T: "Neurogenic potential of Ewing's sarcoma cells." Virchow Archiv. 430 :. 41-46, (1997)

[文献書誌] Ogata,T.: "Impaired male sex development in an infnat with molecularly defined partial 9p monosomy : implication for a testis forming gene (s) on 9p." J Med Genet. 34 :. 331-334, (1997)

[文献書誌] Kikuchi,H.: "Do intronic mutations affecting splicing of WT1 exon 9 cause Frasier syndrome?" J Med Genetics. 35 :. 45-48, (1998)

[文献書誌] Hiraoka,Y.: "Isolation and expression of a human of human SRY-related cDNA,hSOX20." BBA. (in press). (1998)

[文献書誌] Ohkita,H.: "Up-regulated expression of murine EAT,a BCL-2 related gene,in the early stage of differentiaiton of murine embryonal carcinoma cells and embryonic stem cells." BBA. (in press). (1998)

1997 年度実績報告書

秦順一慶應義塾大学, 医学部, 教授 (90051614)