Studies on cytokines and proteases which regulate UV-induced mutation in human cells

调节紫外线诱导的人体细胞突变的细胞因子和蛋白酶的研究

• 批准号: 09680516
• 负责人: SUZUKI Nobuo
• 金额: $ 2.43万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680516/
• 关键词: human cells; ultraviolet ray; mutation; cytokine; protease

In order to clarify molecular mechanisms which regulate mutability in UV(mainly 250nm wavelength) - irradiated human cells, particularly those regulated by proteases and cytokines.1. Partial purification and characterization of UV-induced proteases were preformed and protease samples were obtaind from cell lysates of variant cells with increased levels of the induced protease activity. The samples contained molecules with about 20,000 Mr, the activity of which is inhibited by antipain but enhanced by ALLNa1.2. Using PCR-differential display, we have searched for genes expressed specifically in human interferon (HuIFN)- beta -treated Cockayne syndrome (CS) fibroblast cells. The cDNAs of six clones were similar to expression tagged sequences from unknown genes in databases and the remaining one was identical to the cDNA of the xeroderma pigmentosum XPG gene. Together with our previous finding of increased resistance to ultraviolet (UV) cell-killing of CS cells pretreated with HuIFN- beta … More prior to UV irradiation, suggest that XPG might be one of the genes possibly involved in the HuIFN- beta-induced UV-resistance.3. UV-resistant UV'-10 cells were established from UV-sensitive human RSb cells. We searched here for genes expressed differentially between UV'-10 and RSb cells using a differencial display method to elucidate the molecular mechanisms underlying the UV-resistance. Thirteen candidate cDNA fragment were obtained from 514 mRNA species first screened. Among the cDNA fragments, 3 revealed increased mRNA expression in UV'-10 cells compared with RSb cells by Northern analysis. Nucleotide sequence analysis identified 2 of the 3 cDNA fragments as encoding nucleophosimin (NPM) and T-plastin. The expression of NPM mRNA was induced after UV irradiation in UV'-10 cells but not in RSb cells, whereas irradiation did not affect the expression of T-plastin mRNA.UV'-10 cells transfected with antisense cDNA for NPM mRNA and possibly T-plastin genes may contribute to the incresed resistance to UV cell-killing, at least in the human cells tested. Less

为了阐明紫外线（主要是250 nm波长）照射的人细胞中调节突变的分子机制，特别是蛋白酶和细胞因子调节的机制.对紫外线诱导的蛋白酶进行了部分纯化和表征，并从具有增加的诱导蛋白酶活性水平的变体细胞的细胞裂解物中获得蛋白酶样品。样品含有约20，000 Mr的分子，其活性被抗痛剂抑制，但被ALLNa 1.2增强。使用PCR-差异显示，我们已经搜索了在人干扰素（HuIFN）-β-处理的Cockayne综合征（CS）成纤维细胞中特异性表达的基因。其中6个克隆的cDNA序列与数据库中未知基因的表达标签序列相似，另1个克隆的cDNA序列与着色性干皮病XPG基因的cDNA序列相同。与我们先前发现的用HuIFN-β预处理的CS细胞对紫外线（UV）细胞杀伤的抗性增加一起， ...更多信息 提示XPG基因可能是HuIFN-β诱导的抗紫外基因之一。从紫外线敏感的人RSb细胞建立了抗紫外线的UV '-10细胞。我们在此使用差异显示方法寻找UV-10和RSb细胞之间差异表达的基因，以阐明UV抗性的分子机制。从514种mRNA中筛选出13个候选cDNA片段。在cDNA片段中，北方分析显示3个片段在UV '-10细胞中的mRNA表达比RSb细胞中的mRNA表达高。核苷酸序列分析鉴定了3个cDNA片段中的2个为编码核磷蛋白（NPM）和T-plastin。紫外线照射诱导UV ′-10细胞NPM mRNA的表达，但不诱导RSb细胞的NPM mRNA的表达，而照射不影响T-plastin mRNA的表达。用NPM mRNA和可能的T-plastin基因的反义cDNA转染UV ′-10细胞可能有助于增强对紫外线杀伤的抵抗力，至少在所测试的人细胞中是这样。少

项目成果

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