Hong-zhi Sun1, Lin Xu, Bo Zhou, Wei-jin Zang, Shu-fang Wu. 1. Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, Medical School of Xi'an Jiaotong University, Xi'an 710061, China. sunhongzhi@mail.xjtu.edu.cn
Abstract
AIM: To investigate the role of insulin receptor substrate 2 (IRS-2) in oncogenic transformation induced by v-src. METHODS: IRS-2 gene was silenced using small interfering RNAs (siRNAs). Nuclear translocation and interaction of IRS-2 with v-src was determined using subcellular fractionation, confocal microscopy, and immunoprecipitation. The activity of the cyclin D1 promoter and r-DNA promoter was measured with a luciferase assay. RESULTS: Depletion of IRS-2 inhibited R-/v-src cell growth and reverse the oncogenic transformation. IRS-2 bound to src via its two PI3-K binding sites, which are critical for activities involved in the transformation. Nuclear IRS-2 occupied the cyclin D1 and rDNA promoters. The combination of IRS-2 and v-src increased the activity of the two promoters, especially the rDNA promoter. CONCLUSION: Depletion of insulin receptor substrate 2 could reverse oncogenic transformation induced by v-src.
AIM: To investigate the role of insulin receptor substrate 2 (IRS-2) in oncogenic transformation induced by v-src. METHODS:IRS-2 gene was silenced using small interfering RNAs (siRNAs). Nuclear translocation and interaction of IRS-2 with v-src was determined using subcellular fractionation, confocal microscopy, and immunoprecipitation. The activity of the cyclin D1 promoter and r-DNA promoter was measured with a luciferase assay. RESULTS: Depletion of IRS-2 inhibited R-/v-src cell growth and reverse the oncogenic transformation. IRS-2 bound to src via its two PI3-K binding sites, which are critical for activities involved in the transformation. Nuclear IRS-2 occupied the cyclin D1 and rDNA promoters. The combination of IRS-2 and v-src increased the activity of the two promoters, especially the rDNA promoter. CONCLUSION: Depletion of insulin receptor substrate 2 could reverse oncogenic transformation induced by v-src.
Authors: L Yenush; R Fernandez; M G Myers; T C Grammer; X J Sun; J Blenis; J H Pierce; J Schlessinger; M F White Journal: Mol Cell Biol Date: 1996-05 Impact factor: 4.272