Suppression of decorin expression and partial induction of anchorage-independent growth by the v-src oncogene in human fibroblasts.
Suppression of decorin expression and partial induction of anchorage-independent growth by the v-src oncogene in human fibroblasts.
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v-src 癌基因抑制人成纤维细胞中核心蛋白聚糖的表达并部分诱导不依赖贴壁的生长。
DOI：
10.1046/j.1432-1327.1998.2540266.x
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Robert F. Rosenberger
中科院分区：
文献类型：
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作者：
E. Kolettas;Robert F. Rosenberger
为了确定其在细胞转化中的作用，将 v-src 癌基因引入人胎儿二倍体成纤维细胞 MRC-5 和 MRC-SV1（源自它们的猿猴病毒 40 (SV40) 转化细胞系）中。通过Southern印迹分析确定，感染的细胞含有稳定整合的完整原病毒。尽管高度表达，v-src 并未改变 MRC-5 细胞的形态或生长模式，也未能诱导病灶或改变其饱和密度。然而，v-src 的过度表达降低了 MRC-5 的铺板效率，并在少量但大量的细胞中诱导不依赖贴壁的生长。 Northern印迹分析表明，v-src选择性地消除了核心蛋白聚糖的表达，核心蛋白聚糖是一种小的皮肤素/硫酸软骨素蛋白多糖，与细胞外基质成分相互作用并调节胶原纤维的形成和转化生长因子（TGF）β1的活性。添加除草霉素 A（一种有效的 pp60src 酪氨酸激酶抑制剂）可导致携带 v-src 的 MRC-5 中核心蛋白聚糖的重新表达。纤连蛋白、I 型前胶原或组织纤溶酶原激活剂（一种细胞外基质降解酶的激活剂）的表达没有变化。此外，v-src 不会改变表皮生长因子受体或 TGFbeta1 的表达，也不会减少 MRC-5 成纤维细胞对生长因子的需求。表达v-src的MRC-5和MRC-SV1在注射到裸鼠体内时仍然不致瘤。 v-src 的组成型表达不会改变 c-jun 和 junB 的 mRNA 水平，表明癌基因的作用不是由 AP-1 介导的。先前已证明核心蛋白聚糖基因表达在静止细胞中达到最大，而在转化细胞中几乎不表达。我们的数据表明，在人成纤维细胞中合成核心蛋白聚糖的能力可以被抑制而不发生转化，并且核心蛋白聚糖合成和生长控制之间的关系需要进一步澄清。
To determine its role in cell transformations, the v-src oncogene was introduced into the human foetal diploid fibroblasts MRC-5 and into MRC-SV1, a simian virus 40 (SV40)-transformed cell line derived from them. Infected cells were found to contain stably integrated intact proviruses, as determined by Southern blot analysis. Although highly expressed, v-src did not change the morphology or growth patterns of MRC-5 cells and failed to induce foci or alter their saturation densities. However, overexpression of v-src reduced the plating efficiencies of MRC-5 and induced anchorage-independent growth in a low but significant number of cells. Northern blot analysis showed that v-src selectively abolished the expression of decorin, a small dermatan/chondroitin sulphate proteoglycan that interacts with extracellular-matrix components and modulates collagen-fibril formation and the activity of transforming growth factor (TGF) beta1. Addition of herbimycin A, a potent pp60src tyrosine-kinase inhibitor, resulted in the reexpression of decorin in MRC-5 carrying v-src. There were no changes in the expressions of fibronectin, procollagen type I, or tissue plasminogen activator, an activator of extracellular-matrix-degrading enzymes. Moreover, v-src did not alter the expressions of the epidermal-growth-factor receptor or TGFbeta1 or reduce the growth-factor requirements of MRC-5 fibroblasts. MRC-5 and MRC-SV1 expressing v-src remained non-tumourigenic when injected into nude mice. Constitutive expression of v-src did not alter the mRNA levels of c-jun and junB, suggesting that the effects of the oncogene are not mediated by AP-1. Decorin gene expression has been shown previously to be maximal in quiescent cells and virtually absent in transformed ones. Our data indicate that the ability to synthesise decorin can be suppressed in human fibroblasts without their becoming transformed, and that the relations between decorin synthesis and growth controls need further clarification.