Shen Zhong-Ying, Xu Li-Yan, Li En-Min, Li Jin-Tao, Chen Ming-Hua, Shen Jian, Zeng Yi
Department of Tumor Pathology, Shantou University, Medicine College, Shantou 515031, P.R. China.
Int J Mol Med. 2003 Sep;12(3):341-7.
Ezrin is a key protein in membrane-cytoskeleton interaction. Expression of ezrin in actin-rich cell surfaces may play a role in the modulation of cell shape and adhesion. The aim of this study was to detect ezrin, actin and cytoskeleton and to explore their relationship in the apoptosis of esophageal epithelial cells (SHEE) induced by arsenic trioxide (As2O3). The SHEE is an immortalized human fetal esophageal epithelial cell line, and the cells were treated by administering 5, 10 and 20 micromol of As2O3. the proliferation and apoptosis of SHEE cells were examined by flow cytometry with propidium iodide staining. Ezrin expression was detected by immunocytochemistry and Western blotting. Actin filament was stained by FITC-labeled phalloidin and detected quantitatively by fluorescent microscopy. Cell morphology and microfilaments were examined by electron microscopy. The results revealed that As2O3 induced an inhibition of proliferation and the promotion of apoptosis in SHEE cells. The ezrin, actin and cytoskeleton were decreased after As2O3 treatment and the cellular morphology of apoptosis developed. Our results suggested that the morphological changes of arsenic-induced apoptosis of human esophageal epithelial cells were initiated by ezrin and actin-cytoskeletal aberrance.
埃兹蛋白是膜-细胞骨架相互作用中的关键蛋白。埃兹蛋白在富含肌动蛋白的细胞表面表达可能在细胞形状调节和黏附中发挥作用。本研究旨在检测埃兹蛋白、肌动蛋白和细胞骨架,并探讨它们在三氧化二砷(As2O3)诱导的食管上皮细胞(SHEE)凋亡中的关系。SHEE是一种永生化的人胎儿食管上皮细胞系,细胞分别用5、10和20微摩尔的As2O3处理。采用碘化丙啶染色,通过流式细胞术检测SHEE细胞的增殖和凋亡。通过免疫细胞化学和蛋白质印迹法检测埃兹蛋白表达。用异硫氰酸荧光素标记的鬼笔环肽对肌动蛋白丝进行染色,并通过荧光显微镜进行定量检测。通过电子显微镜检查细胞形态和微丝。结果显示,As2O3诱导SHEE细胞增殖受到抑制,凋亡增加。As2O3处理后,埃兹蛋白、肌动蛋白和细胞骨架减少,细胞凋亡的形态学变化出现。我们的结果表明,砷诱导的人食管上皮细胞凋亡的形态学变化是由埃兹蛋白和肌动蛋白-细胞骨架异常引发的。