The analysis of apoptosis-resistant mechanism in human T lymphocytes

人T淋巴细胞抗凋亡机制分析

• 批准号: 07670208
• 负责人: TAKAHASHI Shuji
• 金额: $ 1.6万
• 依托单位: Sapporo Medical University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670208/
• 关键词: Apoptosis; Fas; glutathione; CD95

Fas antigen is a member of the tumor necrosis receptor family and mediates lethal signal to the Fas-sensitive cells. We previously established the Fas-resistant variant cell lines LAC2D1R and JKT2D1R from the original Fas-sensitive cell lines, SUPT13 and Jurkat, respectively. Recently, we also isolated the Fas-resistant variant CEM2D1R from CCRF-CEM.All of the variants were Fas-positive but resistant to Fas-mediated apoptosis. Further biochemical analysis revealed that the intracellular glutathione (GSH) content of the Fas-resistant variants was higher than in the original cells. When the Fas-resistant variants were incubated with buthionine sulfoximine (BSO) or in GSH-free/cysteine-free medium to deplete GSH,Fas-resistance was reversed. Incubation of the cells with cycloheximide also decreased intracellular GSH and reversed the Fas-resistance. Furthermore, incubation of activated peripheral blood lymphocytes with BSO enhanced Fas-mediated apoptosis. When the Fas-sensitive cells were incubated with n-acetyl cysteine (NAC), intracellular GSH was increased and Fas-mediated apoptosis was blocked. In contrast, Fas-resistant variants, as well as Fas-sensitive cells pretreated with NAC,remained susceptible to allogenic lymphokine-activated killer cells, most likely due to perforin-dependent killing. The results suggest that Fas-mediated apoptosis, but not perforin-dependent killing, is modulated by the intracellular GSH in human T lymphocytes.

Fas抗原是肿瘤坏死受体家族的成员，介导Fas敏感细胞的致死信号。我们先前分别从原始Fas敏感细胞系SUPT 13和Jurkat建立了Fas抗性变体细胞系LAC 2D 1 R和JKT 2D 1 R。最近，我们又从CCRF-CEM中分离到了Fas抗性变异体CEM 2D 1 R，所有变异体均为Fas阳性，但对Fas介导的凋亡具有抗性。进一步的生化分析表明，细胞内谷胱甘肽（GSH）含量的Fas抗性变体高于原始细胞。当Fas抗性变体与丁硫氨酸亚砜（BSO）或在无GSH/无半胱氨酸培养基中孵育以耗尽GSH时，Fas抗性被逆转。放线菌酮孵育的细胞也降低细胞内GSH和逆转Fas抗性。此外，激活的外周血淋巴细胞与BSO孵育增强Fas介导的凋亡。当Fas敏感细胞与N-乙酰半胱氨酸（NAC）孵育时，细胞内GSH增加，Fas介导的凋亡被阻断。相比之下，Fas耐药变异体以及用NAC预处理的Fas敏感细胞仍然对同种异体淋巴因子激活的杀伤细胞敏感，这很可能是由于穿孔素依赖性杀伤。结果表明，Fas介导的细胞凋亡，而不是穿孔素依赖的杀伤，是由人T淋巴细胞内的GSH调制。

项目成果

Takahashi,S.et al: "Immunohistochemical detection of estrogen receptor in invasive human breast cancer:Correlation with heat shock proteins,ps2 and oncogene products" Oncology. 52. 371-375 (1995)

Takahashi,S.et al：“侵袭性人类乳腺癌中雌激素受体的免疫组织化学检测：与热休克蛋白、ps2 和癌基因产物的相关性”肿瘤学。

Inoue, A,Torigoe T,Sogahata K,Kamiguchi K,Takahashi S,Sawada Y,Saijo M,Taya Y,Ishii S,Sato N,Kikuchi K.: "70-kDa heat shock cognate protein interacts directly with the N-terminal region of the retinoblastoma gene product pRb." J Biol Chem. 270. 22571-2257

Inoue, A,Torigoe T,Sogahata K,Kamiguchi K,Takahashi S,Sawada Y,Saijo M,Taya Y,Ishii S,Sato N,Kikuchi K.：“70-kDa 热休克同源蛋白直接与 N 末端相互作用

菊地浩吉,高橋秀史: "アポトーシス-癌とその治療における意義" 医科学出版社(東京), 41 (1996)

Kokichi Kikuchi、Hidefumi Takahashi：“细胞凋亡 - 在癌症及其治疗中的意义”Igaku Shuppansha（东京），41（1996）

Inoue,A.et al: "70-kDa heat shock cognate protein interacts directly with the N-terminal tegion of the retinoblastoma gene product pRb." J.Biol.Chem.270. 22571-22576 (1995)

Inoue,A.等人：“70-kDa 热休克同源蛋白与视网膜母细胞瘤基因产物 pRb 的 N 末端 tegion 直接相互作用。”

Sato,T et al: "Detection of p53 gene mutation on aspiration biopsy specinens from suspected breast cancers by polynerse chain reaction-single strand contormation." Japan J.Cancer.Res.86. 140-145 (1995)

Sato,T 等人：“通过 Polynerse 链式反应 - 单链构造检测疑似乳腺癌的抽吸活检标本中的 p53 基因突变。”