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目的:观察哇巴因对白血病细胞株Jurkat、U937、K562、NB4、HL-60及Raji的增殖和凋亡的影响,并检测哇巴因对Jurkat细胞膜电位的影响。方法:四甲基偶氮唑盐(MTT)法检测哇巴因对不同白血病细胞株生长存活的影响,计算不同浓度哇巴因处理多种白血病细胞株的细胞增殖抑制率;瑞氏染色观察哇巴因对肿瘤细胞生长状态及形态的影响;AnnexinV/PI双染流式细胞仪检测哇巴因分别处理多种细胞的凋亡率;用DiBAC4(3)标记不同浓度哇巴因处理的Jurkat细胞膜电位,流式细胞仪检测细胞膜电位的变化。结果:MTT检测增殖结果表明哇巴因对Jurkat、K562、HL-60、NB4等细胞株均表现出显著增殖抑制作用;细胞形态学观察和凋亡检测结果表明50nmol/L哇巴因处理肿瘤细胞72h大量细胞表现出凋亡的形态特征,Jurkat细胞凋亡率显著增高;DiBAC4(3)标记Jurkat细胞膜电位发现哇巴因没有引起细胞膜的去极化。结论:哇巴因对多种白血病细胞株具有很强的增殖抑制和凋亡诱导作用;哇巴因的增殖抑制和凋亡诱导作用并没有引起Jurkat细胞膜电位的改变,其作用的浓度未引起钠钾ATP酶的抑制。
Objective: To observe the effects of ouabain on the proliferation and apoptosis of leukemia cell lines Jurkat, U937, K562, NB4, HL-60 and Raji and the effect of ouabain on the membrane potential of Jurkat cells. METHODS: The effects of ouabain on the growth and survival of different leukemia cell lines were determined by MTT assay. The cell proliferation inhibition rates of different leukemia cell lines treated with different concentrations of ouabain were calculated. Wright’s staining AnnexinV / PI double staining flow cytometry was used to detect the apoptosis rate of various cells treated with ouabain; Jurkat cell membranes treated with different concentrations of ouabain were labeled with DiBAC4 (3) Potential, flow cytometry to detect changes in cell membrane potential. Results: MTT results showed that ouabain showed significant inhibitory effect on Jurkat, K562, HL-60 and NB4 cell lines. Morphological observation and apoptosis assay showed that 50 nmol / L ouabain could effectively treat tumor cells A large number of cells showed morphological features of apoptosis at 72h, and the apoptosis rate of Jurkat cells was significantly increased. DiBAC4 (3) labeled membrane potential of Jurkat cells found that ouabain did not cause depolarization of cell membrane. CONCLUSION: ouabain has a strong inhibitory effect on proliferation and apoptosis induction in a variety of leukemia cell lines. The inhibition of apoptosis and induction of apoptosis induced by ouabain did not change the membrane potential of Jurkat cells, and its concentration did not induce sodium Potassium ATPase inhibition.