SIMIAN VIRUS-40 LARGE TUMOR-ANTIGEN IS UNABLE TO TRANSFORM MOUSE EMBRYONIC FIBROBLASTS LACKING TYPE-1 INSULIN-LIKE GROWTH-FACTOR RECEPTOR
SIMIAN VIRUS-40 LARGE TUMOR-ANTIGEN IS UNABLE TO TRANSFORM MOUSE EMBRYONIC FIBROBLASTS LACKING TYPE-1 INSULIN-LIKE GROWTH-FACTOR RECEPTOR
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DOI：
10.1073/pnas.90.23.11217
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发表时间：
1993-12-01
影响因子：
11.1
通讯作者：
BASERGA, R
中科院分区：
文献类型：
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作者：
SELL, C;RUBINI, M;BASERGA, R
从编码胰岛素样生长因子I(IGF-I)1型受体(IGF-I)的纯合子小鼠胚胎和它们的野生型窝胎中建立成纤维细胞系。来自野生型胚胎的细胞(W细胞)在添加了血小板衍生生长因子、表皮生长因子和IGF-I的无血清培养液中生长，而来自Igf1r(-/-)胚胎的细胞(R细胞)则不能生长，尽管它们在10%的胎牛血清中生长速度较慢。表达的猴病毒40(SV40)大T抗原可转化W细胞，在单层培养中形成病灶，在软琼脂中形成克隆(非锚定生长)。相比之下，SV40大肿瘤抗原虽然从转基因模板中正常表达，但不能转化R细胞，R细胞仍处于接触抑制状态，无法在软琼脂中生长。如果携带SV40大肿瘤抗原的R细胞稳定地转染表达人IGF-I受体的质粒，转化的表型就会恢复。这些结果表明，通过IGF-I受体的信号转导是SV40转化途径中不可或缺的组成部分。对肿瘤细胞株的反义RNA实验结果进一步支持了这一结论，该结果表明，干扰IGF-I受体的功能对锚定非依赖性生长有深远影响，即使在仅轻微影响单层生长的条件下也是如此。
Fibroblast cell lines were established from mouse embryos homozygous for a targeted disruption of the Igf1r gene, encoding the type 1 receptor for insulin-like growth factor I (IGF-I) and from their wild-type littermates. The cells from the wild-type embryos (W cells) grow in serum-free medium supplemented with platelet-derived growth factor, epidermal growth factor, and IGF-I, whereas the cells from Igf1r(-/-) embryos (R- cells) do not, although they grow at a reduced rate in 10% fetal calf serum. The simian virus 40 (SV40) large T antigen, expressed from a transfected plasmid, can transform W cells, which form foci in monolayer cultures and colonies in soft agar (anchorage-independent growth). In contrast, the SV40 large tumor antigen, although normally expressed from the transfected template, is unable to transform R- cells, which remain contact-inhibited and fail to grow in soft agar. The transformed phenotype is restored if the R- cells carrying the SV40 large tumor antigen are stably transfected with a plasmid expressing the human IGF-I receptor. These results demonstrate that signaling via the IGF-I receptor is an indispensable component of the SV40 transformation pathway. This conclusion is further supported from the results of antisense RNA experiments with tumor cell lines showing that interference with the function of the IGF-I receptor has a profound effect on anchorage-independent growth, even under conditions that only modestly affect growth in monolayers.