Grzanka A, Grzanka D, Zuryń A, Grzanka A A, Safiejko-Mroczka B
Department of Histology and Embrylogy, The Ludwik Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, 85-092 Bydgoszcz, Poland.
Neoplasma. 2006;53(1):56-61.
Influence of taxol, microtubular poison, has been studied on distribution of actin. K-562 and HL-60 cells were treated with taxol in a range of concentrations 0.02-10 microM for 72 hours. The reorganization of F-actin was dependent on its dose. Phalloidin conjugated to TRITC was used to evaluate actin distribution by classical fluorescence and confocal microscopy. Actin was visualized at ultrastructural level by using a postembedding streptavidin-gold method. The treatment of K-562 and HL-60 cells with 2-10 microM of taxol resulted in an increase of F- actin in the cytoplasm, with intense labeling as a ring close to surface of the cell. In HL-60 cells a concentration of F- actin at the site of apoptotic bodies was often observed. Immunogold labeling of actin was localized in the nuclei and cytoplasm in control cells and cells treated with all doses of taxol. At higher doses, compaction of chromatin in the nucleus with strong actin labeling was observed. These observations at the ultrastructural level suggest actin involvement in chromatin reorganization during the process of apoptosis. The present study demonstrated a dose dependent reorganization of actin after treatment with taxol.
已研究了微管毒素紫杉醇对肌动蛋白分布的影响。用浓度范围为0.02 - 10微摩尔的紫杉醇处理K - 562和HL - 60细胞72小时。F - 肌动蛋白的重组取决于其剂量。与TRITC偶联的鬼笔环肽用于通过经典荧光和共聚焦显微镜评估肌动蛋白分布。通过使用包埋后链霉抗生物素蛋白 - 金方法在超微结构水平观察肌动蛋白。用2 - 10微摩尔的紫杉醇处理K - 562和HL - 60细胞导致细胞质中F - 肌动蛋白增加,呈现为靠近细胞表面的环状强烈标记。在HL - 60细胞中,经常观察到凋亡小体部位的F - 肌动蛋白浓度。在对照细胞和用所有剂量紫杉醇处理的细胞中,肌动蛋白的免疫金标记定位于细胞核和细胞质中。在较高剂量下,观察到细胞核中染色质浓缩且伴有强烈的肌动蛋白标记。这些超微结构水平的观察结果表明,在细胞凋亡过程中肌动蛋白参与了染色质重组。本研究证明了紫杉醇处理后肌动蛋白的剂量依赖性重组。