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.

调节人类细胞突变的分子机制是一个有趣的生物学问题。本研究揭示了胰腺癌患者血清因子和人胰腺癌细胞系培养物的条件培养基可增强人RS细胞株的突变性。RS细胞是由人胎儿成纤维细胞建立的，对远紫外光（UV）致突变性高度敏感。与经过照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细胞会导致哇巴因耐药性（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GRP 78/Bip mRNA和蛋白的表达水平降低。使用GRF 78/Bip mRNA的反义寡核苷酸的研究表明，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，通过PCR和差异斑点杂交测定。因此，GRP 78/Bip的下调被认为参与了人细胞突变性的增强。

项目成果

Sugita, T.: "Hypermutable change of human UV^r-1 cells by p53 overexpression"Biochem. Biophys. Res. Commun.. 289. 756-762 (2001)

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。

Nomura, J.: "Increased and decreased expression of CD69 and CD23, respectively, in gravity-stressed lymphocytes"Aviation, Space, and Environmental Medicine. 72. 727-732 (2001)

Nomura, J.：“重力应激淋巴细胞中 CD69 和 CD23 的表达分别增加和减少”航空、航天和环境医学。

Hiwasa, T., et al.: "Increase in ultraviolet sensitivity by overexpression of calpastatin in ultraviolet-resistant UV-1 cells derived from ultraviolet-sensitive human Rsa cells"Cell Death and Differentiation. 7. 531-537 (2000)

Hiwasa, T. 等人：“通过在源自紫外线敏感的人类 Rsa 细胞的抗紫外线 UV-1 细胞中过度表达钙蛋白酶抑制剂来增加紫外线敏感性”细胞死亡和分化。

喜多和子, 他: "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 等人：“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)。