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首页> 外文期刊>Journal of Cellular Physiology >Influence of microenvironment on mammary epithelial cell survival in primary culture.
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Influence of microenvironment on mammary epithelial cell survival in primary culture.

机译：乳腺微环境的影响上皮细胞生存在初级文化。

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摘要

Mammary epithelial cells cultured on Engelbreth-Holm-Swarm (EHS) matrix form multicellular structures termed mammospheres, in which cells and matrix become arranged around a central luminal space. In the presence of lactogenic hormones, cells within mammospheres become polarized, form tight intercellular junctions, and secrete milk proteins vectorially into the luminal space. This study examined the mechanism of lumen formation. Histological examination of developing mammospheres showed that cavitation was associated spatially and temporally with the appearance of fragmented nuclear material in apoptotic bodies, and with the presence of cells positively labeled by terminal deoxynucleotide transferase-mediated deoxyuridine nick end-labeling (TUNEL). Analysis of [(32)P]-deoxynucleotide end-labeled genomic DNA by electrophoresis and autoradiography showed DNA laddering indicative of apoptosis. A transient increase in laddering coincided with both lumen formation and the presence of TUNEL-positive cells. Lumen formation, DNA laddering, and detection of TUNEL-positive cells were all accelerated when matrix composition was altered. They were also impaired coordinately when caspase inhibitor was present during the first two days of culture. Therefore, lumen formation in mammosphere cultures is due to selective apoptosis of centrally located cells. Mammosphere cavitation was accompanied by redistribution of matrix constituents to the mammosphere periphery. Western blotting and Western ligand blotting of culture medium showed that lumen formation was also associated with a transient increase in insulin-like growth factor binding protein-5 (IGFBP5), a factor implicated in mammary apoptosis in vivo. We propose that epithelial cell survival during mammosphere development is induced selectively through stabilization by basement membrane constituents, which may act directly on the epithelial cell or confer protection against autocrine apoptotic factors. Copyright 1999 Wiley-Liss, Inc.
机译：乳腺上皮细胞培养Engelbreth-Holm-Swarm (EHS)矩阵形式多细胞结构称为mammospheres,细胞和矩阵成为周围安排中央腔的空间。在mammospheres刺激乳腺分泌的激素,细胞成为极化,形成紧密的细胞间矢量地连接,分泌的牛奶蛋白质腔的空间。腔的形成机制。检查开发mammospheres显示相关的空间和空化暂时支离破碎的样子在凋亡体核材料,细胞的存在积极的标签终端deoxynucleotide transferase-mediated脱氧尿苷尼克末端标记(TUNEL)。((32) P) -deoxynucleotide end-labeled基因组DNA电泳和放射自显影法显示DNA梯法表明细胞凋亡。瞬态抽丝恰逢的增加腔的形成和存在的TUNEL-positive细胞。抽丝,TUNEL-positive细胞的检测都是加速矩阵组成是什么时候改变。当在半胱天冬酶抑制剂是礼物前两天的文化。形成是由于mammosphere文化选择性集中位于细胞的凋亡。伴随着Mammosphere空化重新分配矩阵组成的mammosphere边缘。西方的培养基的配位印迹显示,内腔的形成也有关瞬态增加胰岛素样生长因子绑定protein-5 (IGFBP5)有牵连的一个因素在乳腺细胞凋亡体内。在mammosphere上皮细胞生存发展是诱导选择性地通过由基底膜成分稳定,这可能直接作用于上皮细胞还是授予保护自分泌凋亡的因素。

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