1996 Fiscal Year Annual Research Report

血管内皮細胞のタイト結合蛋白7H6とがん細胞の血管外脱出の抑制

Research Project

Project/Area Number	08265253
Research Institution	Sapporo Medical University
Principal Investigator	澤田典均札幌医科大学, 医学部, 助教授 (30154149)
Co-Investigator(Kenkyū-buntansha)	小海康夫札幌医科大学, 医学部, 助手 (20178239) 森道夫札幌医科大学, 医学部, 教授 (00045288)
Keywords	タイト結合 / ZO-1 / 7H6(barmotin) / 細胞電気抵抗 / SMC蛋白
Research Abstract	1.目的血管内皮細胞間にはタイト結合が存在し、イオンや蛋白のみならず炎症細胞や癌細胞のintra-およびextra-vasationに関与していると考えられている。したがって、タイト結合の機能(バリア機能)を調節することにより癌細胞の血管への出入りを制御できる可能性がある。我々が発見したタイト結合蛋白のひとつ、7H6蛋白(barmotin)の発現はバリア機能と相関するため、barmotinのcDNA発現クローニング、シークエンスを行い、barmotinの機能を明らかにすると共に、タイト結合の機能調節機構を明かにする。 2.方法と結果 1)株化した肺毛細血管内皮細胞(RLE)をtranswellに単層培養し、その電気抵抗(transendothelial electrical resistance:TER)を測定することにより、タイト結合のバリア機能を評価したところ、db-cAMPとretinoic acidは有意にTERを上昇させるとともに7H6の細胞辺縁への発現を誘導した。この時RLEへの癌細胞のもぐりこみは、対照に比べ約60%抑制された。同様の結果は、腹腔中皮細胞を用いた研究でも得られた。 2)Barmotinは、1184aaからなる蛋白で、既知の蛋白には高いホモロジーを示すものはなかった。構造を調べると、N末とC末にそれぞれWalker's A siteとWalker's B siteと呼ばれるATPaseに特徴的なドメイン構造が離れて存在し、両ドメインの間にはふたつのcoiled coilドメインを有していた。このような構造は最近発見された分子モーターSMC familyに特徴的で、barmotinはSMC1 subfamilyに属することが明らかとなった。

Research Products
(9 results)

All Other

All Publications (9 results)

[Publications] Satoh,H.et al.: "Localization of 7H6 tight junction-associated antigen along the cell border of vascular endothelial cells correlates with paracellular barrier function against ions,large molecules and cancer cells." Exp.Cell.Res.222. 269-274 (1996)
[Publications] Tobioka,H.et al.: "Enhancced paracellular barrier function of rat mesothelial cells partially protects against cacer cell penetration." Brt.J.Cancer. 74. 439-445 (1996)
[Publications] Kimura,M.et al.: "Comparison between the distribution of 7H6 tight junction-associated antigen and occludin during the development of chick intestine." Cell Strut.Funct.21. 91-96 (1996)
[Publications] Kato,J.et al.: "Hepatic iron deprivation prevents spontaneous development of fulminant hepatitis and liver cancer in LEC rats." J.Clin.Invest.98. 923-929 (1996)
[Publications] Obata,H.et al.: "Abnormal accumolation of copper in LEC rat liver induces expression of p53 and nuclear matrix-bound p21^<waf1/cip1>" Carcinogenesis. 17. 2157-2161 (1996)
[Publications] Sawada,N.et al: "Expression of p21^<waf-1/cip-1> is significantly induced in the livers of LEC rats with chronic liver injury." Jpn.J.Cancer Res.87. 1102-1105 (1996)
[Publications] Sawada,N.et al.: "p21^<waf-1/cip-1/sdi-1> is expressed at G1 phase in primary culture of hepatocytes from old rats,presumably prebenting the cells from entering the S phase of the cell cycle." Biochem.Biophys.Res.Commun.228. 819-824 (1996)
[Publications] Kimura,H.et al.: "Bacterial lipopolysaccharide reduced intestinal barrier function and altered localization of 7H6 antigen in IEC-6 rat intestinal crypt cells." J.Cell.Physiol.,. (in press).
[Publications] Yamada-Sasamori,Y.et al: "Rapid induction of 7H6 ti8ght junction-associated protein and paracellular barrier function in capillary endothelial cells of porcine brain in vitro by treatment with astrocyte conditioned medium and cAMP." Med.Electr.Microsc.,. (in press).

1996 Fiscal Year Annual Research Report

血管内皮細胞のタイト結合蛋白7H6とがん細胞の血管外脱出の抑制

Principal Investigator

澤田 典均 札幌医科大学, 医学部, 助教授 (30154149)

Research Products

[Publications] Satoh,H.et al.: "Localization of 7H6 tight junction-associated antigen along the cell border of vascular endothelial cells correlates with paracellular barrier function against ions,large molecules and cancer cells." Exp.Cell.Res.222. 269-274 (1996)

[Publications] Tobioka,H.et al.: "Enhancced paracellular barrier function of rat mesothelial cells partially protects against cacer cell penetration." Brt.J.Cancer. 74. 439-445 (1996)

[Publications] Kimura,M.et al.: "Comparison between the distribution of 7H6 tight junction-associated antigen and occludin during the development of chick intestine." Cell Strut.Funct.21. 91-96 (1996)

[Publications] Kato,J.et al.: "Hepatic iron deprivation prevents spontaneous development of fulminant hepatitis and liver cancer in LEC rats." J.Clin.Invest.98. 923-929 (1996)

[Publications] Obata,H.et al.: "Abnormal accumolation of copper in LEC rat liver induces expression of p53 and nuclear matrix-bound p21^<waf1/cip1>" Carcinogenesis. 17. 2157-2161 (1996)

[Publications] Sawada,N.et al: "Expression of p21^<waf-1/cip-1> is significantly induced in the livers of LEC rats with chronic liver injury." Jpn.J.Cancer Res.87. 1102-1105 (1996)

[Publications] Sawada,N.et al.: "p21^<waf-1/cip-1/sdi-1> is expressed at G1 phase in primary culture of hepatocytes from old rats,presumably prebenting the cells from entering the S phase of the cell cycle." Biochem.Biophys.Res.Commun.228. 819-824 (1996)

[Publications] Kimura,H.et al.: "Bacterial lipopolysaccharide reduced intestinal barrier function and altered localization of 7H6 antigen in IEC-6 rat intestinal crypt cells." J.Cell.Physiol.,. (in press).

[Publications] Yamada-Sasamori,Y.et al: "Rapid induction of 7H6 ti8ght junction-associated protein and paracellular barrier function in capillary endothelial cells of porcine brain in vitro by treatment with astrocyte conditioned medium and cAMP." Med.Electr.Microsc.,. (in press).

澤田典均札幌医科大学, 医学部, 助教授 (30154149)