Zucchi I, Montagna C, Susani L, Vezzoni P, Dulbecco R
Istituto di Tecnologie Biomediche Avanzate, Consiglio Nazionale delle Ricerche, Via Ampere 56, 20131 Milan, Italy.
Proc Natl Acad Sci U S A. 1998 Feb 3;95(3):1079-84. doi: 10.1073/pnas.95.3.1079.
We have developed a model system for studying differentiation in the mammary gland, by using two clonal cultures deriving from a rat breast adenocarcinoma. They differ in the ability to form domes, structures the significance of which is unknown. By using the subtractive cDNA library approach, we isolated a cDNA that is highly expressed in the dome-forming cells, and identical to the rat8 gene and highly homologous to the human 9-27 gene. Antisense treatment of the dome-forming cells specifically and reproducibly abolishes dome formation, while forced expression of the gene in non-dome-forming cells causes morphological changes suggestive of "flat" domes. In situ hybridization on rat tissues shows that the gene is expressed in epithelia, especially in those forming tubular structures, suggesting a relatedness between these structures and domes. Cytokeratin 8 and E cadherin are strongly expressed in the domes but not outside them, suggesting that the rat8 gene triggers the cells to express molecules that tighten the lateral connections between the cells; the process is likely to parallel that occurring during the differentiation of the mammary gland.
我们通过使用源自大鼠乳腺腺癌的两种克隆培养物,开发了一种用于研究乳腺分化的模型系统。它们在形成圆顶结构的能力上有所不同,而圆顶结构的意义尚不清楚。通过使用消减cDNA文库方法,我们分离出一种在形成圆顶的细胞中高度表达的cDNA,它与大鼠8基因相同,并且与人类9 - 27基因高度同源。对形成圆顶的细胞进行反义处理可特异性且可重复地消除圆顶的形成,而在不形成圆顶的细胞中强制表达该基因会导致形态变化,提示形成“扁平”圆顶。对大鼠组织进行原位杂交显示该基因在上皮细胞中表达,特别是在形成管状结构的上皮细胞中,这表明这些结构与圆顶之间存在相关性。细胞角蛋白8和E - 钙黏蛋白在圆顶中强烈表达,但在圆顶外不表达,这表明大鼠8基因触发细胞表达能够加强细胞间侧向连接的分子;这一过程可能与乳腺分化过程中发生的过程相似。