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目的 研究氨基葡萄糖硫酸盐对白血病细胞K5 6 2增殖的影响 ,并探讨分子机制。方法 采用MTT(四唑氮蓝 )法检测不同浓度的氨基葡萄糖硫酸盐对K5 6 2细胞生长的影响 ;吉姆萨 -瑞氏染色、电镜观察药物作用后形态的改变 ;流式细胞仪检测药物对K5 6 2细胞细胞周期的影响和细胞表面分化抗原CD11b、CD14、CD33和CD13的表达变化 ;WesternBlot检测药物作用前后K5 6 2细胞组织蛋白酶D(CathepsinD)表达的改变。结果 氨基葡萄糖硫酸盐能明显抑制K5 6 2细胞的生长 ,且存在剂量依赖性。与未加药的对照相比 ,在 5mmol·L-1浓度作用 2d后形态学观察 ,易见早、中、晚期凋亡的K5 6 2细胞 ;胞质空泡化明显 ,大泡很多。 1、5mmol·L-1浓度作用 2d后出现G1期K5 6 2细胞增加 ,而S期的减少 ,在 5mmol·L-1浓度时 ,出现凋亡峰与对照相比 ,CD11b和CD14的表达轻度增加 ,CD33表达轻度减低 ,而CD13表达无明显变化。同时还发现药物作用后出现CathepsinD蛋白前体的蓄积现象。结论 氨基葡萄糖硫酸盐能抑制K5 6 2细胞增殖 ,诱导其凋亡 ,其分子机制可能与细胞周期受阻、CathepsinD蛋白前体蓄积有关
Objective To study the effect of glucosamine sulfate on the proliferation of leukemia K562 cells and to explore the molecular mechanism. Methods MTT (tetrazolium blue) was used to detect the effects of different concentrations of glucosamine sulfate on the growth of K5 6 2 cells. The changes of morphological changes were observed with Giemsa-Wright staining and electron microscopy. K5 6 2 cells and the expression of cell surface differentiation antigens CD11b, CD14, CD33 and CD13. Western Blot was used to detect the expression of Cathepsin D in K562 cells before and after drug treatment. Results Glucosamine sulfate significantly inhibited the growth of K5 6 2 cells in a dose-dependent manner. Compared with the untreated control, K562 cells which were early, middle and late apoptotic cells were easily observed after treated with 5mmol·L-1 for 2 days. The cytoplasmic vacuoles were obvious and many big bubbles appeared. After treated with 1,5 mmol·L-1 for 2 days, K5 6 2 cells increased in G1 phase and decreased in S phase. The apoptotic peak appeared at 5 mmol·L-1 concentration, and the expression of CD11b and CD14 was lighter Degree increased, CD33 expression decreased slightly, while CD13 expression had no significant change. The accumulation of Cathepsin D protein precursor was also observed after drug action. Conclusion Glucosamine sulfate inhibits the proliferation and induces apoptosis of K562 cells. The molecular mechanism may be related to the cell cycle arrest and Cathepsin D protein precursor accumulation