Wall Michelle E, Rachlin Andrew, Otey Carol A, Loboa Elizabeth G
Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, North Carolina 27695-7115, USA.
Am J Physiol Cell Physiol. 2007 Nov;293(5):C1532-8. doi: 10.1152/ajpcell.00065.2007. Epub 2007 Aug 8.
Cell morphology may be an important stimulus during differentiation of human adipose-derived adult stem (hADAS) cells, but there are limited studies that have investigated the role of the cytoskeleton or associated proteins in hADAS cells undergoing differentiation. Palladin is an actin-associated protein that plays an integral role in focal adhesion and cytoskeleton organization. In this study we show that palladin was expressed by hADAS cells and was modulated during osteogenic differentiation and in response to cyclic tensile strain. Human ADAS cells expressed the 90- and 140-kDa palladin isoforms and upregulated expression of both isoforms after culture in conditions that promoted osteogenesis. Palladin mRNA expression levels were also increased in hADAS cells subjected to cyclic tensile strain. Knockdown of the palladin gene during osteogenesis resulted in decreased actin stress fibers and decreased protein levels of Eps8, an epidermal growth factor receptor tyrosine kinase that colocalizes with actin. Silencing the palladin gene, however, did not affect hADAS cells' commitment down the osteogenic lineage.
细胞形态可能是人类脂肪来源的成体干细胞(hADAS细胞)分化过程中的一个重要刺激因素,但研究细胞骨架或相关蛋白在hADAS细胞分化过程中作用的研究有限。Palladin是一种与肌动蛋白相关的蛋白质,在粘着斑和细胞骨架组织中发挥着不可或缺的作用。在本研究中,我们发现hADAS细胞表达Palladin,并且在成骨分化过程中以及对周期性拉伸应变的反应中其表达受到调控。人ADAS细胞表达90 kDa和140 kDa的Palladin异构体,在促进成骨的条件下培养后,两种异构体的表达均上调。在经受周期性拉伸应变的hADAS细胞中,Palladin mRNA表达水平也增加。在成骨过程中敲低Palladin基因导致肌动蛋白应力纤维减少,以及与肌动蛋白共定位的表皮生长因子受体酪氨酸激酶Eps8的蛋白水平降低。然而,沉默Palladin基因并不影响hADAS细胞向成骨谱系的分化。