注射用核糖核酸Ⅱ上调p53诱导人白血病细胞凋亡

Ribonucleic acid Ⅱ induces apoptosis in human leukemia cells by up-regulating p53

目的研究注射用核糖核酸Ⅱ诱导人白血病细胞系K562和KG1a细胞凋亡的作用。方法以K562和KG1a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;Hoechst33258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、200mg·L^(-1)注射用核糖核酸Ⅱ处理K562和KG1a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;Hoechst33258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KG1a细胞凋亡。

Aim To investigate the effect of ribonucleic acid Ⅱon apoptosis in human leukemia cell lines K562 and KG1a. Methods Cell counting kit-8（ CCK-8） assay 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. Expression levels of p53,Bax,Bcl-2 and cleaved caspase-3were analyzed by Western blot. Results The proliferation 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 indicated the inhibitory effect of ribonucleic acid Ⅱ was in dose-dependent and time-dependent manners. FCM results displayed a dose-dependent increase in cell apoptotic 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. Western 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 ribonucleic 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.