Studies on suppression of transformation in primary rat embryo fibroblasts

抑制原代大鼠胚胎成纤维细胞转化的研究

• 批准号: 07670244
• 负责人: INOUE Hirokazu
• 金额: $ 1.47万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670244/
• 关键词: Transformation; Oncogene; Tumor suppressor gene; Primary cells; Colon cancer; がん抑制遺伝子; 癌化抑制

On searching for transformation suppressor genes in primary cells, we isolated a novel gene, drs (doenregulated by v-src), which was expressed in normal rat fibroblast cells but completely suppressed in the cells transformed by the v-src gene, from a cDNA library of REF.The molecular cloned cDNA was about 1.8kb in size, containing an open reading frame composed of 464 amino acid residues. Homology search of drs protein revealed that this protein had one transmembrane domain and three consensus repeats (CR) which were conserved as various numbers of CR in extracellular domain of the selsctin family. Expression of drs mRNA was also down-regulated by serum stimulation of G0-arrested normal rat cells. To clarify the biological function of the drs gene, we introduced the drs gene linked to potent promoter into normal rat cell line and examined the sensitivity to transformation by the v-src oncogene. Rat cell lines expressing exogenous drs gene showed lower efficiency of v-src transformation, indicating a transformation suppressor function of the drs gene. The drs gene was also downregulated in human colon cancer cell lines and ectopic expression of this gene suppressed the ability of prolifelation in these cancer cell lines, suggesting a role in the genesis of human cancers.

在原代细胞中寻找转化抑制基因的过程中，我们从REF的cDNA文库中分离到一个新基因drs（doenregulated by v-src），该基因在正常大鼠成纤维细胞中表达，而在v-src基因转化的细胞中完全被抑制。同源性分析表明，drs蛋白具有1个跨膜区和3个保守重复序列（consensus repeats，CR），在selsctin家族的胞外区具有不同数量的CR。G 0期阻滞的正常大鼠细胞的血清刺激也下调drs mRNA的表达。为了阐明drs基因的生物学功能，我们将drs基因与有效启动子连接，导入正常大鼠细胞系，并检测v-src癌基因转化的敏感性。表达外源drs基因的大鼠细胞系v-src转化效率较低，表明drs基因具有转化抑制功能。drs基因在人结肠癌细胞系中也下调，并且该基因的异位表达抑制了这些癌细胞系中的增殖能力，表明其在人类癌症发生中的作用。

项目成果

Tavoloni, N.and Inoue, H.: "Cellular aging is a critical determinant of primary cell resistance to v-Src transformation." J.Virol.71. 237-247 (1997)

Tavoloni, N. 和 Inoue, H.：“细胞衰老是原代细胞抵抗 v-Src 转化的关键决定因素。”

N.Tavoloni and H.Inoue: "Cellular aging is a critical determinant of primary cell resistance to v-Src transformation." J.Virol.71 1. 237-247 (1997)

N.Tavoloni 和 H.Inoue：“细胞衰老是原代细胞抵抗 v-Src 转化的关键决定因素。”

G.Tang et al.: "A mutant cell line resistant to Vibrioparahaemolyticus termostulde direct hemolysin (TDH) : its potential in identification of putative receptor of TDH." Biochem.Biophys.Acta. (in press). (1997)

G.Tang 等人：“对副溶血弧菌直接溶血素 (TDH) 具有抗性的突变细胞系：其在识别假定的 TDH 受体方面的潜力。”

M.Shimakage et al.: "Tumorigenicity of EBNA2-transfected cells." FEBS Letters. 371. 245-248 (1995)

M.Shimakage 等人：“EBNA2 转染细胞的致瘤性”。

M.Shimakage et al.: "Tumorigenicity of EBNA2-transfected cells" FEBS letters. 371. 245-248 (1995)

M.Shimakage 等人：“EBNA2 转染细胞的致瘤性”FEBS 字母。