Molecular mechanisms of mutation enhancement by serum factors from pancreatic cancer patients and purification of the factors.

胰腺癌患者血清因子增强突变的分子机制及其纯化。

• 批准号: 12671141
• 负责人: KITA Kazuko
• 金额: $ 2.11万
• 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671141/
• 关键词: PANCREATIC CANCER; K-; RAS CODON 12; GENE MUTATION; GENE EXPRESSION; MOLECUALR CHAPERONE; HUMAN CELLS

The molecular mechanisms that regulate mutability of human cells are intriguing biological problems. The present study revealed that mutability of human RS cell strain was enhanced by serum factors From pancreatic cancer patients and conditioned medium from cultures of a human pancreatic cancer cell line. RS cells, established from human fetal fibroblasts, are highly sensitive to far-rultraviolet light (UV) mutagenicity. Preculture of human RS cells with the serum factors or the medium prior to irradiation with UV resulted in an increase in the frequency of ouabain resistance (Oua^R) mutations, compared with cells irradiated but not treated with either the serum factors or the medium. Decreases in expression levels of the molecular chaperone GRP78/Bip mRNA and protein were observed in the medium-treated RSa cells. Studies using antisense oligonucleotides for GRF78/Bip mRNA showed that the antisense oligonucieotide-treated RSb cells were more susceptible to UV-induced K-ras codon 12 base substitution mutations than mock-treated cells, as determined by PCR and differential dot-blot hybridization. Thus, down-regulation of GRP78/Bip was suggested to be involved in the enhancement of mutability of human cells.

调节人类细胞突变性的分子机制是令人感兴趣的生物学问题。本研究表明，来自胰腺癌患者的血清因子和来自人胰腺癌细胞系培养物的条件培养基增强了人RS细胞株的突变性。 RS 细胞由人类胎儿成纤维细胞建立，对远紫外光 (UV) 致突变性高度敏感。与经辐射但未用血清因子或培养基处理的细胞相比，在用紫外线照射之前用血清因子或培养基预培养人RS细胞导致哇巴因抗性(Oua^R)突变频率增加。在培养基处理的 RSa 细胞中观察到分子伴侣 GRP78/Bip mRNA 和蛋白质表达水平下降。使用反义寡核苷酸检测 GRF78/Bip mRNA 的研究表明，通过 PCR 和差异点印迹杂交测定，反义寡核苷酸处理的 RSb 细胞比模拟处理的细胞更容易受到紫外线诱导的 K-ras 密码子 12 碱基取代突变的影响。因此，GRP78/Bip 的下调被认为与人类细胞突变性的增强有关。

项目成果

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Sugita, T.：“p53 过表达导致人类 UV^r-1 细胞发生超突变”Biochem。

Moriya, T.: "Enhanced expression of the LDH-A gene after gravity-changing stress in human RSa cells"Biol. Sci. Space. (in press).

Moriya, T.：“人类 RSa 细胞中重力变化应激后 LDH-A 基因的表达增强”Biol。

喜多和子: "自由落下による微小重力環境を利用したヒト細胞の変異誘導調節機能に関する研究"J. The Japan Society of Microgravity Application. 18. 180-184 (2001)

Kazuko Kita：“利用自由落体引起的微重力环境对人体细胞突变诱导的调节研究” J. 日本微重力应用学会 18. 180-184 (2001)。

Takahashi, S.: "Mutagenicity of bisphenol A and its suppression by interferon-α in human RSa cells"Mutat. Res.. 490. 199-207 (2001)

Takahashi, S.：“人 RSa 细胞中双酚 A 的致突变性及其受干扰素 α 的抑制”Res.. 490. 199-207 (2001)

Takahashi, S.: "A novel mehtod of analyzing environmental chemical mutagens in human cell systems"Water science and Technology. 4288(7-8). 133-138 (2000)

Takahashi, S.：“一种分析人体细胞系统中环境化学诱变剂的新方法”水科学与技术。