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组织蛋白酶(cathepsin)D在氨基葡萄糖硫酸盐(glucosaminesulfate,GS)诱导的慢性髓系白血病K562细胞凋亡过程中为激活线粒体途径的可能的中介分子。为了探讨BclxL在氨基葡萄糖硫酸盐诱导慢性髓系白血病K562细胞凋亡中的可能作用,在倒置显微镜下动态观察细胞生长变化,采用瑞氏-姬姆萨染色观察细胞加药前后的形态学改变,间接免疫荧光法观察GS诱导K562细胞凋亡过程中cathepsinD和细胞色素(cytochrome)C的转位情况,Westernblot检测K562细胞凋亡过程中BclxL、Bax、Bid蛋白的表达变化。结果表明:5mmol/L氨基葡萄糖硫酸盐作用于K562细胞72小时后,细胞增殖受到明显抑制,胞浆出现空泡化;荧光显微镜显示cathepsinD和cytochromeC的荧光信号由颗粒状趋向弥散;BclxL蛋白在GS加药组中表达减低,甚至消失,但在抑胃酶肽(pepstatin)A预处理后的加药组中表达没有明显改变,Bax和Bid蛋白表达在凋亡前后没有明显变化。结论:BclxL蛋白在GS诱导白血病K562细胞凋亡过程中可能介导cathepsinD与线粒体途径之间的联系。
Cathepsin D is a potential mediator of mitochondrial pathway during the apoptosis of chronic myeloid leukemia K562 cells induced by glucosaminesulfate (GS). In order to investigate the possible role of BclxL in the apoptosis of K562 cells induced by glucosamine sulphate, the growth of K562 cells was observed under an inverted microscope. The morphological changes of cells were observed by Wright-Giemsa staining. Indirect immunofluorescence method was used to observe the translocation of cathepsinD and cytochrome C during the apoptosis of K562 cells induced by GS. The expressions of BclxL, Bax and Bid proteins in K562 cells were detected by Western blot. The results showed that the proliferation of K562 cells was inhibited by 5 mmol / L glucosamine sulfate for 72 hours, and the cytoplasm was vacuolized. Fluorescence microscopy showed that the fluorescence signals of cathepsin D and cytochrome C tended to diffuse from granule. The expression of BclxL protein in GS However, the expression of Bax and Bid did not change significantly before and after apoptosis in the dosing group after pepstatin A treatment. CONCLUSION: BclxL protein may mediate the link between cathepsin D and mitochondrial pathway in the process of GS-induced leukemia K562 cell apoptosis.