初代培養肝細胞の温熱耐性と熱ショック蛋白72の局在に関する研究
初代培養肝細胞を用いて温熱刺激(heat shock: HS)時のstress fibers (SF), cell viability (CV), heat shock protein 72 (HSP 72)について検討した.SFは43℃, 45℃のHSにてCVの低下に先行して迅速に消失し,軽度のHSでは可逆的に回復した.SFの消失は可逆性の早期の細胞障害の指標となり得ると考えた.CVはMTT法とmethylene blue法で測定したが,HS時のCV測定値はMTT法の方が著明な変化を示した.初代培養肝細胞の温熱耐性は43℃ 20分間のHSによって時間とともに獲得され,8時間から12時間で最大...
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| Published in | 肝臓 Vol. 32; no. 5; pp. 505 - 511 |
|---|---|
| Main Authors | , , , , , , , |
| Format | Journal Article |
| Language | Japanese |
| Published |
一般社団法人 日本肝臓学会
1991
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0451-4203 1881-3593 |
| DOI | 10.2957/kanzo.32.505 |
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| Abstract | 初代培養肝細胞を用いて温熱刺激(heat shock: HS)時のstress fibers (SF), cell viability (CV), heat shock protein 72 (HSP 72)について検討した.SFは43℃, 45℃のHSにてCVの低下に先行して迅速に消失し,軽度のHSでは可逆的に回復した.SFの消失は可逆性の早期の細胞障害の指標となり得ると考えた.CVはMTT法とmethylene blue法で測定したが,HS時のCV測定値はMTT法の方が著明な変化を示した.初代培養肝細胞の温熱耐性は43℃ 20分間のHSによって時間とともに獲得され,8時間から12時間で最大であり,それには核小体へ集積するHSP 72が重要な役割を果たしていることが示唆された.これらの成績は,癌温熱療法の治療成績の向上に役立つものと考えられる. |
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| AbstractList | 初代培養肝細胞を用いて温熱刺激(heat shock: HS)時のstress fibers (SF), cell viability (CV), heat shock protein 72 (HSP 72)について検討した.SFは43℃, 45℃のHSにてCVの低下に先行して迅速に消失し,軽度のHSでは可逆的に回復した.SFの消失は可逆性の早期の細胞障害の指標となり得ると考えた.CVはMTT法とmethylene blue法で測定したが,HS時のCV測定値はMTT法の方が著明な変化を示した.初代培養肝細胞の温熱耐性は43℃ 20分間のHSによって時間とともに獲得され,8時間から12時間で最大であり,それには核小体へ集積するHSP 72が重要な役割を果たしていることが示唆された.これらの成績は,癌温熱療法の治療成績の向上に役立つものと考えられる. |
| Author | 岡上, 武 加嶋, 敬 香川, 恵造 今西, 二郎 森本, 道雄 長尾, 泰孝 山口, 希 伊藤, 義人 |
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| References | 9) Glass JR, Dewitt RG, Cress AE: Rapid loss of stress fibers in Chinese hamster overy cells after hyperthermia. Cancer Res 45: 258-262, 1985 16) 藤原敬己:ストレスファイバーの形態と機能.蛋核酵 34: 1513-1523, 1989 17) Schamhart DHJ, Boonstra J, Graft MV, et al: The occurrence of thermotolerance in non-proliferating rat hepatocytes in primary culture. Int J Radiat Biol 47: 213-218, 1985 2) Mizzen LA, Welch WJ: Characterization of the thermotolerant cell. II. Effects on the intracellular distribution of heat-shock protein, intermediate filaments, and small nuclear ribonucleoprotein complexes. J Cell Biol 106: 1117-1130, 1988 1) Mizzen LA, Welch WJ: Characterization of the thermotolerant cell. I. Effects on protein synthesis activity and the regulation of heatshock protein 70 expression. J Cell Biol 106: 1105-1116, 1988 5) Welch WJ: 高等動物ストレス(熱ショック)蛋白質の構造と機能.実験医学 7: 1989-2000, 1989 14) Iida K, Iida H, Yahara I: Heat shock induction of intranuclear actin rods in cultured mammalian cells. Exp Cell Res 165: 207-215, 1986 4) Welch WJ, Suhan JP: Cellular and biochemical events in mammalian cells during and after recovery from physiological stress. J Cell Biol 103: 2035-2052, 1986 10) Carmicheal J, DeGraff WG, Gazdar AF, et al: Evaluation of a tetrazolium-based semiautomated colorimetric assay: Assessment of chemosensitivity testing. Cancer Res 47: 936-942, 1987 13) Finlay GJ, Baguley BC, Wilson WR: A semiautomated microculture method for investigated growth inhibitory effects of cytotoxic compounds on exponentially growing carcinoma cells. Anal Biochem 139: 272-277, 1984 11) Mosmann T: Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immun Methods 65: 55-63, 1983 3) Riabowol KT, Mizzen LA, Welch WJ: Heat shock is lethal to fibroblasts microinjected with antibodies against hsp 70. Science 242: 433-436, 1988 18) Landry J, Chretien P, Bernier D, et al: Thermotolerance and heat shock proteins induced by hyperthermia in rat liver cells. Int J Radiat Oncol Biol Phys 8: 59-62, 1982 15) 永森静志,藤瀬清隆,蓮村哲,他:培養肝細胞を用いた障害実験.肝胆膵 19: 269-275, 1989 12) Yamazaki S, Onishi E, Enami K, et al: Proposal of standardized methods and reference for assaying recombinant human tumor necrosis factor. Jap J Med Sci Biol 39: 105-118, 1986 7) Ichihara A, Nakamura T, Tanaka K: Use of hepatocytes in primary culture for biochemical studies on liver functions. Mol Cell Biochem 43: 145-162, 1982 8) Seglen PO: Preparation of isolated rat liver cell. Methods Cell Biol 13: 29-83, 1976 6) Landry J, Bernier D, Chretien P, et al: Synthesis and degradation of heat shock proteins during development and decay of thermotolerance. Cancer Res 42: 2457-2461, 1982 |
| References_xml | – reference: 7) Ichihara A, Nakamura T, Tanaka K: Use of hepatocytes in primary culture for biochemical studies on liver functions. Mol Cell Biochem 43: 145-162, 1982 – reference: 2) Mizzen LA, Welch WJ: Characterization of the thermotolerant cell. II. Effects on the intracellular distribution of heat-shock protein, intermediate filaments, and small nuclear ribonucleoprotein complexes. J Cell Biol 106: 1117-1130, 1988 – reference: 5) Welch WJ: 高等動物ストレス(熱ショック)蛋白質の構造と機能.実験医学 7: 1989-2000, 1989 – reference: 15) 永森静志,藤瀬清隆,蓮村哲,他:培養肝細胞を用いた障害実験.肝胆膵 19: 269-275, 1989 – reference: 3) Riabowol KT, Mizzen LA, Welch WJ: Heat shock is lethal to fibroblasts microinjected with antibodies against hsp 70. Science 242: 433-436, 1988 – reference: 6) Landry J, Bernier D, Chretien P, et al: Synthesis and degradation of heat shock proteins during development and decay of thermotolerance. Cancer Res 42: 2457-2461, 1982 – reference: 12) Yamazaki S, Onishi E, Enami K, et al: Proposal of standardized methods and reference for assaying recombinant human tumor necrosis factor. Jap J Med Sci Biol 39: 105-118, 1986 – reference: 14) Iida K, Iida H, Yahara I: Heat shock induction of intranuclear actin rods in cultured mammalian cells. Exp Cell Res 165: 207-215, 1986 – reference: 18) Landry J, Chretien P, Bernier D, et al: Thermotolerance and heat shock proteins induced by hyperthermia in rat liver cells. Int J Radiat Oncol Biol Phys 8: 59-62, 1982 – reference: 1) Mizzen LA, Welch WJ: Characterization of the thermotolerant cell. I. Effects on protein synthesis activity and the regulation of heatshock protein 70 expression. J Cell Biol 106: 1105-1116, 1988 – reference: 10) Carmicheal J, DeGraff WG, Gazdar AF, et al: Evaluation of a tetrazolium-based semiautomated colorimetric assay: Assessment of chemosensitivity testing. Cancer Res 47: 936-942, 1987 – reference: 13) Finlay GJ, Baguley BC, Wilson WR: A semiautomated microculture method for investigated growth inhibitory effects of cytotoxic compounds on exponentially growing carcinoma cells. Anal Biochem 139: 272-277, 1984 – reference: 8) Seglen PO: Preparation of isolated rat liver cell. Methods Cell Biol 13: 29-83, 1976 – reference: 11) Mosmann T: Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immun Methods 65: 55-63, 1983 – reference: 4) Welch WJ, Suhan JP: Cellular and biochemical events in mammalian cells during and after recovery from physiological stress. J Cell Biol 103: 2035-2052, 1986 – reference: 16) 藤原敬己:ストレスファイバーの形態と機能.蛋核酵 34: 1513-1523, 1989 – reference: 9) Glass JR, Dewitt RG, Cress AE: Rapid loss of stress fibers in Chinese hamster overy cells after hyperthermia. Cancer Res 45: 258-262, 1985 – reference: 17) Schamhart DHJ, Boonstra J, Graft MV, et al: The occurrence of thermotolerance in non-proliferating rat hepatocytes in primary culture. Int J Radiat Biol 47: 213-218, 1985 |
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| Snippet | 初代培養肝細胞を用いて温熱刺激(heat shock: HS)時のstress fibers (SF), cell viability (CV), heat shock protein 72 (HSP 72)について検討した.SFは43℃,... |
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| Title | 初代培養肝細胞の温熱耐性と熱ショック蛋白72の局在に関する研究 |
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