Expression of a down-regulated target, SSeCKS, reverses v-Jun-induced transformation of 10T1/2 murine fibroblasts
Expression of a down-regulated target, SSeCKS, reverses v-Jun-induced transformation of 10T1/2 murine fibroblasts
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DOI：
10.1038/sj.onc.1204077
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发表时间：
2001-01-11
期刊：
影响因子：
8
通讯作者：
Vogt, PK
中科院分区：
文献类型：
--
作者：
Cohen, SB;Waha, A;Vogt, PK
过表达v-Jun癌蛋白的10 T1/2小鼠成纤维细胞系在培养中仅发生形态学改变，生长成多层病灶，悬浮于琼脂中形成集落。v-Jun转化的10 T1/2细胞的生长速率与未转化的亲本细胞相比无显著变化，但转化培养物的饱和密度超过正常对照的2倍，从v-Jun转化的10 T1/2细胞中提取mRNA，用DNA微阵列技术分析差异基因表达。v-Jun下调的靶点之一被鉴定为SSeCKS（Src抑制的C激酶底物）。SSeCKS在v-Jun转化的成纤维细胞中的再表达逆转了细胞的转化表型。它们形成病灶的能力降低到背景水平，琼脂菌落的数量和大小降低了10倍，饱和密度显著降低。然而，SSeCKS的表达对v-Jun转化的10 T1/2细胞的形态几乎没有影响。这些数据表明SSeCKS蛋白具有生长衰减特性。SSeCKS的下调可能是Jun-induced转化所必需的。
Line 10T1/2 mouse fibroblast overexpressing the v-Jun oncoprotein mere morphologically altered, grew into multilayered foci in culture and formed colonies when suspended in agar, The growth rate of the v-Jun-transformed 10T1/2 cells was not changed significantly from that of the untransformed parental cells, but the saturation density of the transformed cultures exceeded that of normal controls by a factor of 2, mRNA extracted from v-Jun-transformed 10T1/2 cells was analysed for differential gene expression with DNA micro-array technology. One of the targets downregulated by v-Jun was identified as SSeCKS (Src-suppressed C kinase substrate). Re-expression of SSeCKS in v-Jun-transformed fibroblasts reversed the transformed phenotype of the cells. Their ability to form foci was reduced to background levels, the number and size of agar colonies was lowered by a factor of 10 and the saturation density was significantly diminished. However, expression of SSeCKS had little effect on the morphology of v-Jun-transformed 10T1/2 cells. These data suggest that the SSeCKS protein has growth-attenuating properties. Down-regulation of SSeCKS may be essential for Jun-induced transformation.