Michalczyk A, Brown R W, Collins J P, Ackland M L
Deakin University, Centre for Cellular and Molecular Biology, School of Biological and Chemical Sciences, Burwood Campus, Burwood, Victoria 3125, Australia.
Differentiation. 2001 Feb;67(1-2):41-9. doi: 10.1046/j.1432-0436.2001.067001041.x.
The human breast contains two epithelial lineages, luminal epithelial and myoepithelial. Specific patterns of expression of intermediate filaments have previously been demonstrated in the resting breast. To determine how terminal differentiation and lactation influenced expression of intermediate filaments in breast epithelial cells, we used Western blot analysis to measure the levels of vimentin, alpha-smooth muscle actin, keratin 14, and keratin 18 in the resting and lactating breast. Confocal immunofluorescence was used to determine the subcellular site of localization of the intermediate filaments. Vimentin was localised to myoepithelial cells in both the resting and lactating gland. There was a four-fold increase in vimentin protein levels in lactating tissue relative to resting tissue, and this may be related to increased cellular activity of the myoepithelial cells which surround secretory alveoli. Alpha-smooth muscle actin and keratin 14 were detected in myoepithelial cells, and similar levels of expression were found in lactating and resting tissue. In the resting breast, keratin 18 and keratin 8 were detected in luminal epithelial cells in a filamentous form, whereas in lactating tissue it was present in a punctate form in luminal cells and also seen as granules in the lumen of alveoli. Our results indicate that intermediate filament expression patterns are altered in the lactating human breast, and this may reflect their role in the fully functional gland.
人类乳腺包含两种上皮谱系,即管腔上皮和肌上皮。此前已在静止乳腺中证实了中间丝的特定表达模式。为了确定终末分化和泌乳如何影响乳腺上皮细胞中中间丝的表达,我们使用蛋白质免疫印迹分析来测量静止期和泌乳期乳腺中波形蛋白、α-平滑肌肌动蛋白、角蛋白14和角蛋白18的水平。共聚焦免疫荧光用于确定中间丝的亚细胞定位位点。波形蛋白在静止期和泌乳期腺体的肌上皮细胞中均有定位。相对于静止期组织,泌乳期组织中波形蛋白的蛋白质水平增加了四倍,这可能与围绕分泌性肺泡的肌上皮细胞的细胞活性增加有关。在肌上皮细胞中检测到α-平滑肌肌动蛋白和角蛋白14,在泌乳期和静止期组织中发现了相似的表达水平。在静止期乳腺中,角蛋白18和角蛋白8在管腔上皮细胞中以丝状形式被检测到,而在泌乳期组织中,它在管腔细胞中以点状形式存在,并且在肺泡腔中也可见为颗粒状。我们的结果表明,泌乳期人类乳腺中中间丝的表达模式发生了改变,这可能反映了它们在功能完整的腺体中的作用。