1992 Fiscal Year Annual Research Report

ラット胸腺腫発生遺伝子Tsr-1の作用機序に関する研究

Research Project

Project/Area Number	04151026
Research Institution	Nagoya University
Principal Investigator	松山睦司名古屋大学, 医学部, 教授 (80073112)
Co-Investigator(Kenkyū-buntansha)	羽藤文彦大阪市立大学, 医学部, 講師 (00198772) 増田彰愛知県がんセンター研究所, 研究員 (50157202) 松浦晃洋札幌医科大学, 医学部, 講師 (70157238) 木下喜博大阪市立大学, 医学部, 教授 (80046896) 田口修愛知県がんセンター研究所, 主任研究員 (00142167)
Keywords	BUF / Mna系ラット / 胸腺腫 / 胸腺増大遺伝子 / Ten-1 / 胸腺腫発生遺伝子 / Tsr-1 / TGF-β / EGFR
Research Abstract	1.X因子の精製:ラット胸腺上皮細胞株IT-45R1をEarle's塩類溶液にて24時間培養後,培養上清を逆相カートリッジにて濃縮し、陰イオン交換カラムおよびゲル濾過カラムを用いて分画し、培養ラット胸腺リンパ球の[^3H]thymidineの取り込みに与える影響を検討したところ、分子量340-150kDaの分画にトリプシン感受性で比較的熱耐性のリンパ球増殖誘導因子が分離された。 2.Tsr-1遺伝子導入コンジェニックラットの樹立:Tsr-1遺伝子を胸腺腫嫌発系のACI/NMs系ラットに導入すべく退交配を繰り返し行い、10代に達した。 3.胸腺腫上皮細胞が産生するマクロファージ活性化因子の解析:(1)胸腺腫上皮細胞株TaD1-3およびTa5-4の培養上清には、胸腺マクロファージの増殖・融合を促進する活性、および正常胸腺初代培養における上皮細胞の増殖を促進する活性が存在した。(2)この活性はゲル濾過カラムによって約100倍濃縮され、56℃30分処理で安定、99℃5分処理またはトリプシンによって失活することがわかった。(3)マクロファージの融合と活性化の程度と胸腺および胸腺腫の重量との間に顕著な正の相関が観察された。 4.胸腺腫発生における上皮細胞・リンパ球の相互作用に関する検討:胎齢15日のBUF/Mna系ラットより胸腺を採取し、BALB/cヌードおよびscidマウスの腎被膜下に移植して、胸腺腫発生の有無を観察した。ヌードマウスには臓器局在性自己免疫病が、scidマウスにはGVH様自己免疫病の発症が認められた。致死量の放射線を照謝した胎仔胸腺をscidマウスに移植した場合には、上皮細胞のみの生着が認められた。 5.Tsr-1遺伝子の分子生物学的検索:(1)胸腺増大遺伝子Ten-1、胸腺腫感受性遺伝子Tsr-1、およびラットヌード遺伝子rnuのassignment:{(WKY X BUF/Mna)F1 X BUF/Mna}、{ACI/NMs X (BUF/Mna X ACI/NMs)F1}、および{BUF/Mna-rnu/rnu X WHY)F1 X WHY}退交配ラットを用いて、RFLP法およびPCR法による多数のprobeの系統間多型を指標として、Ten-1、Tsr-1およびrnuの染色体との連関を検索した。Ten-1およびTsr-1はNo.1、rnuはNo.10染色体上に位置することが判明した。(2)正常胸腺、胸腺腫および悪性胸腺腫由来の合計7種のin vitro培養株細胞におけるproto-oncogeneおよびtumor suppressor gene合計15種につきNorthern blot解析を行ったところ、悪性胸腺腫由来細胞において著明なTGF-β発現増加とEGFR発現低下が見い出された。(3)胸腺腫発生に到る種々の時期におけるリンホカイン遺伝子発現の変動をRT-PCR法により検討したところ、BUF/Mna系のラットのみに特異的に、IL-4が胸腺および胸腺腫のサイズが一定に達した12週および72週以降で発現の増加がみられた。

Research Products

(22 results)

All Other

All Publications (22 results)

[Publications] Taguchi,O.: "An intrinsic thymic epithelial abnormality is responsible for the spontaneous development of predominantly lymphocytic thymomas in BUF/Mna rats." Jpn.J.Cancer Res.83. 1166-1171 (1992)
[Publications] Masuda,A.: "Abnormal increase in multinuclear macrophages in primary cultures of BUF/Mna rat thymomas." Virchows Arch.[B]Cell Path.62. 125-132 (1992)
[Publications] Lu,J.: "Establishment of transplantable tumor lines and in vitro cell lines of malignant thymoma developed in a BUF/Mna rat." Jpn.J.Cancer Res.83. 618-624 (1992)
[Publications] Sakai,Y.: "Expression of proto-oncogenes and tumor suppressor genes in in vitro cell lines derived from a thymus,thymoma,and malignant thymoma of rats." Nagoya J.Med.Sci.55. (1993)
[Publications] Kinoshita,Y.: "Target cells for new thymocyte growth factor derived from the rat thymus epithelial cell line(TAD3)." Recent Advances in Cellular and Molecular Biology. 1. 19-24 (1992)
[Publications] Hato,F.: "Preparation of monoclonal antibody against synthetic thymosin α_1 and its effect on the biological activity of thymosin α_1f." Recent Advances in Cellular and Molecular Biology. 1. 35-41 (1992)
[Publications] Tominaga,K.: "Enhancement of DNA synthesis in rat thymocytes by stimulating their muscarinic acetylcholine receptors." Cell.Mol.Biol.38. 815-822 (1992)
[Publications] Wajjwalku,W.: "A gene encoding ligand for deletion of T cells bearing TcrVβ6 and Vβ8.1 cosegregates with new endogenous mouse mammary tumor virus." Immunogenet.(1993)
[Publications] Masuda,A.: "Signal transduction cascade shared by epidermal growth factor and platelet-derived growth factor is a major pathway for oncogenic transformation in NRK cells." New Biologist. 4. 489-503 (1992)
[Publications] Taniguchi,M.: "Establishment and characterization of a malignant melanocytic tumor cell lines expressing the ret oncogene." Oncogene. 7. 1491-1496 (1992)
[Publications] Wajjwalku,W.: "Low frequency of the ret and trk proto-oncogenes in Japanese thyroid papillary carcinomas." Jpn.J.Cancer Res.83. 671-675 (1992)
[Publications] Isomura,T.: "RFP is a DNA binding protein associated with the nuclear matrix." Nucl.Acid.Res.20. 5305-5310 (1992)
[Publications] Kawasaki,S.: "Polyoxyethylene-modified superoxide dismutase reduces side effects of adriamycin and mitomycin C." Jpn.J.Cancer Res.83. 899-906 (1992)
[Publications] Takayama,S.: "Molecular genetic analysis of class Ib MHC-related genes.Identification of rat homologues of Tla and CD1." Sapporo Med.J.61. 49-60 (1992)
[Publications] Murakami,T.: "Structure and expression of the rat homologue of CD5." Sapporo Med.J.61. 13-26 (1992)
[Publications] Kojima,A.: "Inhibitory effect by 1,25-dihydroxyvitamin D_3 on concanavalin A-stimulated proliferation of rat thymic lymphocytes." Cell.Mol.Biol.38. 867-875 (1992)
[Publications] Sakakibara,Y.: "Development of thermotolerance in hsp70 induction-defective mutant of NRK cells." Int.J.Hyperthermia. 8. 329-340 (1992)
[Publications] Kontani,K.: "Involvement of the H^+/K^+-ATPase α subunit as a major antigenic protein in autoimmune gastritis induced by neonatal thymectomy in mice." Clin.Exp.Immunol.89. 63-67 (1992)
[Publications] Miyaishi,O.: "Distribution of the collagen binding hear-shock protein in chicken tissues." J.Histochem.Cytochem.40. 1021-1029 (1992)
[Publications] Torii,K.: "Aging-associated deletions of human diaphramatic mitochondrial DNA." Am.J.Respir.Cell Mol.Biol.6. 543-549 (1992)
[Publications] Ikeda,H.: "Analysis of T cell receptor V region gene usage of cytotoxic T lymphocytes and tumor infiltration lymphocytes derived from human autologous gastric signet ring cell carcinomas." Cancer Res.
[Publications] Uesugi,Y.: "Effects of sex steroids on the development of sexual dimorphism in mouse innominate hone." Anat.Rec.243. 541-548 (1992)

1992 Fiscal Year Annual Research Report

ラット胸腺腫発生遺伝子Tsr-1の作用機序に関する研究

Principal Investigator

松山 睦司 名古屋大学, 医学部, 教授 (80073112)

Research Products

[Publications] Taguchi,O.: "An intrinsic thymic epithelial abnormality is responsible for the spontaneous development of predominantly lymphocytic thymomas in BUF/Mna rats." Jpn.J.Cancer Res.83. 1166-1171 (1992)

[Publications] Masuda,A.: "Abnormal increase in multinuclear macrophages in primary cultures of BUF/Mna rat thymomas." Virchows Arch.[B]Cell Path.62. 125-132 (1992)

[Publications] Lu,J.: "Establishment of transplantable tumor lines and in vitro cell lines of malignant thymoma developed in a BUF/Mna rat." Jpn.J.Cancer Res.83. 618-624 (1992)

[Publications] Sakai,Y.: "Expression of proto-oncogenes and tumor suppressor genes in in vitro cell lines derived from a thymus,thymoma,and malignant thymoma of rats." Nagoya J.Med.Sci.55. (1993)

[Publications] Kinoshita,Y.: "Target cells for new thymocyte growth factor derived from the rat thymus epithelial cell line(TAD3)." Recent Advances in Cellular and Molecular Biology. 1. 19-24 (1992)

[Publications] Hato,F.: "Preparation of monoclonal antibody against synthetic thymosin α_1 and its effect on the biological activity of thymosin α_1f." Recent Advances in Cellular and Molecular Biology. 1. 35-41 (1992)

[Publications] Tominaga,K.: "Enhancement of DNA synthesis in rat thymocytes by stimulating their muscarinic acetylcholine receptors." Cell.Mol.Biol.38. 815-822 (1992)

[Publications] Wajjwalku,W.: "A gene encoding ligand for deletion of T cells bearing TcrVβ6 and Vβ8.1 cosegregates with new endogenous mouse mammary tumor virus." Immunogenet.(1993)

[Publications] Masuda,A.: "Signal transduction cascade shared by epidermal growth factor and platelet-derived growth factor is a major pathway for oncogenic transformation in NRK cells." New Biologist. 4. 489-503 (1992)

[Publications] Taniguchi,M.: "Establishment and characterization of a malignant melanocytic tumor cell lines expressing the ret oncogene." Oncogene. 7. 1491-1496 (1992)

[Publications] Wajjwalku,W.: "Low frequency of the ret and trk proto-oncogenes in Japanese thyroid papillary carcinomas." Jpn.J.Cancer Res.83. 671-675 (1992)

[Publications] Isomura,T.: "RFP is a DNA binding protein associated with the nuclear matrix." Nucl.Acid.Res.20. 5305-5310 (1992)

[Publications] Kawasaki,S.: "Polyoxyethylene-modified superoxide dismutase reduces side effects of adriamycin and mitomycin C." Jpn.J.Cancer Res.83. 899-906 (1992)

[Publications] Takayama,S.: "Molecular genetic analysis of class Ib MHC-related genes.Identification of rat homologues of Tla and CD1." Sapporo Med.J.61. 49-60 (1992)

[Publications] Murakami,T.: "Structure and expression of the rat homologue of CD5." Sapporo Med.J.61. 13-26 (1992)

[Publications] Kojima,A.: "Inhibitory effect by 1,25-dihydroxyvitamin D_3 on concanavalin A-stimulated proliferation of rat thymic lymphocytes." Cell.Mol.Biol.38. 867-875 (1992)

[Publications] Sakakibara,Y.: "Development of thermotolerance in hsp70 induction-defective mutant of NRK cells." Int.J.Hyperthermia. 8. 329-340 (1992)

[Publications] Kontani,K.: "Involvement of the H^+/K^+-ATPase α subunit as a major antigenic protein in autoimmune gastritis induced by neonatal thymectomy in mice." Clin.Exp.Immunol.89. 63-67 (1992)

[Publications] Miyaishi,O.: "Distribution of the collagen binding hear-shock protein in chicken tissues." J.Histochem.Cytochem.40. 1021-1029 (1992)

[Publications] Torii,K.: "Aging-associated deletions of human diaphramatic mitochondrial DNA." Am.J.Respir.Cell Mol.Biol.6. 543-549 (1992)

[Publications] Ikeda,H.: "Analysis of T cell receptor V region gene usage of cytotoxic T lymphocytes and tumor infiltration lymphocytes derived from human autologous gastric signet ring cell carcinomas." Cancer Res.

[Publications] Uesugi,Y.: "Effects of sex steroids on the development of sexual dimorphism in mouse innominate hone." Anat.Rec.243. 541-548 (1992)

松山睦司名古屋大学, 医学部, 教授 (80073112)