Characterization of the Wilms tumor gene product WT1

维尔姆斯肿瘤基因产物 WT1 的表征

• 批准号: 04454171
• 负责人: AKIYAMA Tetsu
• 金额: $ 3.9万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454171/
• 关键词: Transcription factor; Wilms tumor; Tumor suppressor gene; WT1; cell cycle; differentiation; cyclin; CDK; leukemia; Zinc finger; 増殖; zinc finger; splicing; WT遺伝子; WT蛋白質

We examined the function of WT1 in cellular proliferation and differentiation, and obtained the following results.1. Microinjection of the WT1 cDNA in NIH3T3 cells blocked serum-induced cell cycle progression into S phase. The inhibitory activity of WT1 was abrogated by the overexpression of cyclin E/CDK2 as well as cyclin D1/CDK4. Furthermore, both CDK4- and CDK2-associated kinase activities were downregulated in cells overexpressing WT1, whereas the levels of CDK4, CDK2 and cyclin D1 expression were unchanged. These findings suggest that inhibition of the activity of cyclin/CDK complexes may be involved in mediating the WT1-induced cell cycle block.2. The expression of WT1 decreased during retinoic acid-induced differentiation of embryonal carcinoma F9 cells. Cell lines constitutively expressing the exogenous WT1 failed to differentiate into primitive endoderm cells and underwent apoptotic death, suggesting that downregulation of WT1 expression is necessary for normal differentiation of F9 cells into parietal endoderm cells. We also found that WT1 is a new prognostic factor and a new marker for the detection of minimal residual disease in acute leukeia.3. Using a yeast two-hybrid system, we obtained a novel cDNA clone which encodes a putative WT1-associated protein. Detailed characterization of this protein is underway in our laboratory.

本研究检测了WT 1在细胞增殖和分化中的作用，获得以下结果.显微注射WT 1 cDNA可阻断血清诱导的细胞周期进入S期。WT 1的抑制活性被细胞周期蛋白E/CDK 2和细胞周期蛋白D1/CDK 4的过表达所废除。此外，CDK 4和CDK 2相关激酶活性在过表达WT 1的细胞中下调，而CDK 4、CDK 2和细胞周期蛋白D1表达水平不变。这些发现表明，抑制细胞周期蛋白/CDK复合物的活性可能参与介导WT 1诱导的细胞周期阻滞。2.在维甲酸诱导胚胎癌F9细胞分化过程中，WT 1的表达下降。组成型表达外源性WT 1的细胞系未能分化成原始内胚层细胞并经历凋亡性死亡，这表明WT 1表达的下调对于F9细胞正常分化成壁内胚层细胞是必要的。我们还发现WT 1是一个新的预后因子，也是检测急性白血病微小残留病的一个新的标志物.利用酵母双杂交系统，我们获得了一个新的cDNA克隆，它编码一个推定的WT 1相关蛋白。我们的实验室正在对这种蛋白质进行详细的表征。

项目成果

T.Kudoh et al.: "G1 phase arrest induced by Wilms tumor protein WT1 is abrogated by cyclin/CDK." Proc.Natl.Sci.USA.92. 4517-4521 (1995)

T.Kudoh 等人：“Wilms 肿瘤蛋白 WT1 诱导的 G1 期阻滞被细胞周期蛋白/CDK 消除。”

K.Inoue, H.Sugiyama, H.Ogawa, M.Nakagawa, T.Yamagami, H.Miwa, K.Kita, A.Hiraoka, T.Masaoka, K.Nasu, T.Kyo, H.Dohy, H.Nakauchi, T.Ishidate, K.Akiyama and T.Kishimoto: "WT1 as a new prognostic factor and a new marker for the detection of minimal residual di

K.Inoue、H.Sugiyama、H.Okawa、M.Nakakawa、T.Yamagami、H.Miwa、K.Kita、A.Hiraoka、T.Masaoka、K.Nasu、T.Kyo、H.Dohy、H.

T.Kudoh, T.Ishidate, M.Moriyama, K.Toyoshima and T.Akiyama: "G1 phase arrest induced by Wilms tumor protein WT1 is abrogated by cyclin/CDK." Proc.Natl.Acad.Sci.USA. 92. 4517-4521 (1995)

T.Kudoh、T.Ishidate、M.Moriyama、K.Toyoshima 和 T.Akiyama：“Wilms 肿瘤蛋白 WT1 诱导的 G1 期阻滞被细胞周期蛋白/CDK 消除。”

T.Akiyama: "Transcription factor as tumor suppressor gene product." Exp.Med. (Jikken Igaku). 11. 191-196 (1993)

T.Akiyama：“作为抑癌基因产物的转录因子。”

門脇孝: "細胞増殖因子の最前線" 羊土社, 182 (1993)

Takashi Kadowaki：“细胞生长因子的最前沿”Yodosha，182（1993）