AMPHIREGULIN AS AN AUTOCRINE GROWTH-FACTOR FOR C-HA-RAS-TRANSFORMED AND C-ERBB-2-TRANSFORMED HUMAN MAMMARY EPITHELIAL-CELLS
AMPHIREGULIN AS AN AUTOCRINE GROWTH-FACTOR FOR C-HA-RAS-TRANSFORMED AND C-ERBB-2-TRANSFORMED HUMAN MAMMARY EPITHELIAL-CELLS
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DOI：
10.1073/pnas.91.7.2790
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发表时间：
1994-03-29
影响因子：
11.1
通讯作者：
SALOMON, DS
中科院分区：
文献类型：
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作者：
NORMANNO, N;SELVAM, MP;SALOMON, DS
双调蛋白（AR）是表皮生长因子（EGF）家族的一员，在刺激永生化、非转化的人乳腺上皮MCF-10A细胞的贴壁依赖性生长（ADG）方面与EGF一样有效。与MCF-10A细胞相比，通过活化的人c-Ha-ras原癌基因（MCF-10A ras）或通过过表达未活化的大鼠c-neu基因（MCF-10A neu）转化的MCF-10A细胞在ADG中对AR的反应降低了35%，但在这些转化体中AR仍与EGF一样有效。外源性AR在刺激癌基因转化的MCF-10A细胞的锚定非依赖性生长（一种通常依赖于外源性EGF的反应）方面仅表现出EGF活性的15-20%。MCF-10A细胞表达低水平的1.4-kb AR mRNA转录物，而MCF-10A ras和MCF-10A neu细胞显示AR mRNA和内源性AR蛋白水平增加15- 30倍，如通过Western印迹分析测定的。外源性EGF可诱导MCF-10A亲本细胞和转化细胞中AR mRNA和蛋白的表达。与AR mRNA的5'端序列互补的20-mer硫代磷酸反义脱氧寡核苷酸能够显著降低内源性AR蛋白的水平，并抑制EGF刺激的MCF-10A ras和MCF-10A neu细胞的ADG和锚定非依赖性生长。这些数据表明，AR可能作为一个EGF依赖性自分泌生长因子在乳腺上皮细胞已被转化的点突变的c-Ha-ras或c-neu。
Amphiregulin (AR), a member of the epidermal growth factor (EGF) family, was found to be as potent as EGF in stimulating the anchorage-dependent growth (ADG) of immortalized, nontransformed human mammary epithelial MCF-10A cells. MCF-10A cells transformed by either an activated human c-Ha-ras protooncogene (MCF-10A ras) or by overexpression of a nonactivated rat c-neu gene (MCF-10A neu) exhibited a 35% reduction in the response to AR in ADG when compared to MCF-10A cells, but AR was still as potent as EGF in these transformants. Exogenous AR exhibited only 15-20% of the activity of EGF in stimulating the anchorage-independent growth, a response that is normally dependent upon exogenous EGF, of the oncogene-transformed MCF-10A cells. MCF-10A cells express low levels of a 1.4-kb AR mRNA transcript, while MCF-10A ras and MCF-10A neu cells display a 15- to 30-fold increase in the levels of AR mRNA and endogenous AR protein as determined by Western blot analysis. Exogenous EGF was found to induce both AR mRNA and protein in the MCF-10A parental and transformed cells. A 20-mer phosphorothioate antisense deoxyoligonucleotide complementary to the 5' sequence of AR mRNA was able to significantly reduce the levels of endogenous AR protein and to inhibit the EGF-stimulated ADG and anchorage-independent growth of MCF-10A ras and MCF-10A neu cells. These data suggest that AR may function as an EGF-dependent autocrine growth factor in mammary epithelial cells that have been transformed by either a point-mutated c-Ha-ras or c-neu.