Development of yeast-based screening method for cancer-related genes and its clinical applications

基于酵母的癌症相关基因筛选方法的建立及其临床应用

• 批准号: 09557206
• 负责人: ISHIOKA Chikashi
• 金额: $ 6.4万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 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-09557206/
• 关键词: genetic diagnosis; functional analysis; Saccharomyces cerevisiae

We have developed the yeast-based diagnostic system for cancer susceptibility genes, including the BRCA 1, BRCA2, ATM, APC, hMSH2 and hMLH 1 genes. Among these, former five genes were adapted to the yeast-based stop codon assay (SC assay) which has the ability to detect nonsense and frameshift mutations within the open reading frame of the genes. Only in case of hMLH I gene, a functional assay, which has the ability to detect and characterize the missense mutations, was developed. Since germline missense mutations have been often reported in hereditary non-polyposis colorectal cancer families. the assay we developed here should be extremely useful for detection and characterization of pathogenic mutations of the hMLH1 gene. We are successfully improving these assay systems by using green fluorescence protein (GFP) gene as a marker. The resulting assays became to be powerful genetic diagnostic tools for detection of cancer susceptibility genes.

我们开发了基于酵母的癌症易感基因诊断系统，包括BRCA 1、BRCA2、ATM、APC、hMSH2和hmlh1基因。其中，前5个基因适用于基于酵母的停止密码子试验（SC试验），该试验具有检测基因开放阅读框内的无义和移码突变的能力。仅在hMLH I基因的情况下，开发了一种功能分析，具有检测和表征错义突变的能力。由于种系错义突变在遗传性非息肉病性结直肠癌家族中经常被报道。我们在这里开发的检测方法对于检测和表征hMLH1基因的致病性突变非常有用。我们利用绿色荧光蛋白（GFP）基因作为标记，成功地改进了这些检测系统。由此产生的检测方法成为检测癌症易感基因的强有力的遗传诊断工具。

项目成果

石岡千加史 他: "新培養細胞実験法 (黒木登志夫ら編)" 羊土社, 316 (1999)

Chikashi Ishioka 等：“新培养细胞实验方法（Toshio Kuroki 等编辑）” Yodosha，316（1999）

Chikashi Ishioka: "Oligomerization is not esential for growth suppression by p53 in p53-deficient osteosarcoma Saos-2 cells." Biochem.Biophys.Res.Commun.232. 54-60 (1997)

Chikashi Ishioka：“在 p53 缺陷的骨肉瘤 Saos-2 细胞中，寡聚化对于 p53 生长抑制并不是必需的。”

Ishioka, C.: "Detection of heterogygous truncating mutations in the BRCA1 and APC genes by using a rapid screening assay in yeast." Proc.Natl.Acad.Sci., USA. 94. 2449-2553 (1997)

Ishioka, C.：“通过在酵母中使用快速筛选测定来检测 BRCA1 和 APC 基因中的异型截短突变。”

Hrainer, M.: "Differential contributions of BRCA1 and BRCA2 to early-onset hreastcancer." N.Enge.J.Med.336. 1416-1421 (1997)

Hrainer, M.：“BRCA1 和 BRCA2 对早发性乳腺癌的不同贡献。”

Li-Qun Jia: "Screening the p53 status of human cell lines using a yeast functional assay." Mol.Carcinog.19. 243-253 (1997)

Li-Qun Jia：“使用酵母功能测定筛选人类细胞系的 p53 状态。”