注射用核糖核酸Ⅱ上调p53诱导人白血病细胞凋亡

摘要

目的：研究注射用核糖核酸Ⅱ诱导人白血病细胞系K562和KG1 a细胞凋亡的作用。方法以K562和KG1 a细胞为研究对象，采用CCK-8法检测注射用核糖核酸Ⅱ对细胞增殖的影响；流式细胞术检测细胞凋亡率的变化；Hoechst 33258染色观察细胞核形态；免疫印迹技术检测p53及凋亡相关蛋白Bcl-2、Bax和cleaved caspase-3的表达。结果以100～300 mg·L－1注射用核糖核酸Ⅱ处理K562和KG1a细胞12、24、48 h后，CCK-8检测结果显示K562和KG1a细胞的增殖降低，抑制率明显增高，并呈时间剂量依赖性；100、150、200 mg·L－1注射用核糖核酸Ⅱ处理K562和KG1 a细胞24 h后， FCM结果显示细胞凋亡率随药物剂量增加而增高；Hoechst 33258染色观察到细胞核出现染色质浓缩聚集、染色加深等凋亡指征；Western blot结果显示，注射用核糖核酸Ⅱ上调p53、Bax和cleaved caspase-3的表达，下调Bcl-2表达。结论注射用核糖核酸Ⅱ通过上调p53，调控Bcl-2／Bax的表达，激活caspase-3诱导人白血病K562和KG1 a细胞凋亡。%Aim To investigate the effect of ribonucleic acidⅡon apoptosis in human leukemia cell lines K562 and KG1 a.Methods Cell counting kit-8(CCK-8)as-say was performed to detect proliferation activity of K562 and KG1 a cells treated with ribonucleic acidⅡ. Apoptosis index was assessed by flow cytometry(FCM) and fluorescent Hoechst 33258 staining was used for observing morphologic changes of apoptosis.Expres-sion levels of p53,Bax,Bcl-2 and cleaved caspase-3 were analyzed by Western blot.Results The prolifera-tion of K562 and KG1 a cells was significantly inhibited by ribonucleic acid Ⅱ treatment for 12 h,24 h,48 h at concentrations of 100~300 mg·L-1 ,which indica-ted the inhibitory effect of ribonucleic acid Ⅱ was in dose-dependent and time-dependent manners.FCM re-sults displayed a dose-dependent increase in cell apop-totic rate.Hoechst 33258 staining showed the typical apoptotic morphology in some leukemic cells treated with ribonucleic acid Ⅱ,including increased nuclear chromatin concentration and edge accumulation.West-ern blot analysis showed the increased expression of p53,Bax,cleaved caspase-3 and decreased expression of Bcl-2 in K562 and KG1 a cells treated with ribonu-cleic acid Ⅱ.Conclusions Ribonucleic acid Ⅱ can induce apoptosis of leukemia K562 and KG1 a cells by up-regulating p53,which mediates Bcl-2/Bax balance and activates caspase-3 .