Study on the interleukin-1 signaling

IL-1信号传导研究

• 批准号: 06672194
• 负责人: ONOZAKI Kikuo
• 金额: $ 1.34万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06672194/
• 关键词: Interleukin 1; interleukin 1 receptor; cell growth; signal transduction; polyamine; ODC; インターロイギン1; レセプター; サイトカイン

The mechanisms of IL-latiproliferative effect and the acquired resistance to IL-1 effect were studied using human melanoma cells A375. As type I IL-1 receptor (IL-1R) transduces IL-1 signal, the regulatory mechanisms of IL-1R expression in vitro and in vivo were also studied.1. The mechanism of IL-1 antiproliferative effectIL-1R cDNA was transfected into A375-5 which is resistant to IL-1 because it expresses very low number of IL-1R.We obtained the transfectant which became sensitive to IL-1. Using this clone we studied the IL-1 signaling. IL-1 downregulated ODC activity. Although ODC mRNA level was not changed, the amount of protein of ODC decreased. The mRNA of antizyme (AZ), a proteinous factor which inactivates ODC by binding to ODC and causes degradation of ODC,was upregulated by IL-1. Furthermore, antisense of AZ inhibited IL-1 effects. Terefore, AZ induced by IL-1 appeared to be important in the IL-1 signaling.2. The mechanism of acquired resistance to IL-1Using the cells which acquired resistance to IL-1 antiproliferative effect after long period of culture, we studied the mechanism. Expression vectors of IL-1alpha sense and IL-1alpha antisense were transfected into IL-1 sensitive and resistant cells, respectively. However, the sensitivity of the transfectants to IL-1 was not changed. Therefore, expression of IL-1alpha appeared not to be sufficient to the resistance.3. Regulation of IL-1R expressionUsing human fibroblast cell line TIG-1, we studied the regulatory mechanism of type I IL-1R wxpression. Tyrosine kinase appeared to play an important role in constitutive expression of IL-1R.Whtn mice were injected with LPS,a marked increase of IL-1R mRNA was induced in the liver. The upregulation apperaed to be induced by the endogenously produced IL-1 and IL-6.

以人黑色素瘤细胞A375为研究对象，研究了IL-1促增殖作用和获得性抵抗IL-1作用的机制。由于IL-1受体(IL-1R)是IL-1信号的转导通路，本研究还对IL-1R在体内外表达的调控机制进行了研究。IL-1R的抗增殖作用机制将IL-1R基因导入对IL-1产生抗性的A375-5细胞，获得对IL-1敏感的A375-5。利用该克隆，我们研究了IL-1信号转导途径。IL-1下调ODC活性。虽然ODC的mRNA水平没有变化，但ODC的蛋白质含量减少。抗酶(AZ)是一种蛋白因子，通过与ODC结合而使ODC失活，导致ODC降解，IL-1上调了AZ的表达。此外，反义AZ可抑制IL-1的作用。因此，IL-1诱导的AZ似乎在IL-1信号转导中起重要作用。IL-1获得性耐药的机制利用长期培养获得IL-1耐药细胞的抗增殖作用，探讨其机制。IL-1α正义表达载体和IL-1α反义表达载体分别导入IL-1敏感和耐药细胞。但转基因细胞对IL-1的敏感性没有改变。因此，IL-1α的表达似乎不足以抵抗。利用人成纤维细胞系TIG-1，研究了I型IL-1R表达的调控机制。酪氨酸激酶在IL-1R的组成性表达中起重要作用。这种上调可能是由内源性产生的IL-1和IL-6诱导的。

项目成果

D. Yang, T. Takii, H. Hayashi S. Ito, M. Hayashi & K. Onozaki: "Molecular cloning of human antizyml cDNA" Biochemistry and Molecular Biology International. (in press). (1996)

D. Yang、T. Takii、H. Hayashi S. Ito、M. Hayashi

S.Ito, H.Hayashi, N.Watanabe, Y.Kobayashi, T.Takii and K.Onozaki: "Interleukin 1 (IL-1) production is not essential for acquired resistance of human melanoma cells A375 to anti-proliferative effect of IL-1." Int.J.Cancer. (in press). (1996)

S.Ito、H.Hayashi、N.Watanabe、Y.Kobayashi、T.Takii 和 K.Onozaki：“白细胞介素 1 (IL-1) 的产生对于人黑色素瘤细胞 A375 的抗增殖作用的获得性抵抗并不重要。

Y.Takii,K.Wada,T.Chiba,H.Hayashi,T.Ishihara,K.Onzaki: "Development of glycosylated human IL-1α,neoglyco IL-1α,coupled caith D-mannose demier;Synthesis and biologcal activities in vitro." LYMPHOKINE AND CYTOKINE RESEARCH. 13. 265-270 (1994)

Y. Takii、K. Wada、T. Chiba、H. Hayashi、T. Ishihara、K. Onzaki：“糖基化人 IL-1α、新糖 IL-1α、偶联 caith D-甘露糖二聚体的开发；合成和生物活性淋巴细胞因子和细胞因子研究 13. 265-270 (1994)

K.Wada,T.Chiba,Y.Takei,H.Ishihara,H.Hayashi,K.Onzaki: "Synthes is of immunosuppressive neoglycoproteins:Bovine serum albumin coupled with 8(hydrazino-carbnoyl)octyl4-or6-0-α-D.Man-α-D-Man." Journal of CARBOHYDRATE CHEMISTRY. 13. 941-965 (1994)

K. Wada、T. Chiba、Y. Takei、H. Ishihara、H. Hayashi、K. Onzaki：“合成免疫抑制性新糖蛋白：牛血清白蛋白与 8（肼基-羰酰基）辛基 4-或 6-0-α- 偶联D.Man-α-D-Man。”《碳水化合物化学杂志》。13. 941-965 (1994)

T. Takii, H. Hayashi T. Marunouchi & K. Onozaki: "Interleukin 1 down rogulates type I interleukin 1 mRNA exprossion in a human fibroblast cell line TIG-1 in the absence of prostaglandin E_2 synthesis." Lymphokine and Cytokine Research. 13. 213-219 (1994)

T.泷井、H.林 T.丸之内