桃儿七可促进慢性粒细胞白血病K562细胞的凋亡

目的 探讨藏药桃儿七对慢性粒细胞白血病K562细胞凋亡的影响及相关机制.方法 用不同浓度的桃儿七对K562细胞株进行不同时间梯度的处理,应用CCK8试验筛选桃儿七的最佳作用浓度及时间;流式细胞术检测细胞凋亡;瑞氏染色观察细胞形态学改变,DAPI染色观察细胞核形态变化;Western blotting分别检测凋亡相关蛋白PARP、caspase-3、Cleaved-caspase3的活化情况以及BCR/ABL、p-BCR/ABL、STAT5、p-STAT5蛋白表达变化.结果 分别以1、2、3μg/mL浓度的桃儿七处理细胞12、24、36、48、72、96 h,K562细胞增殖呈浓度时间依赖性抑制,并筛选出2μg/mL,48 h为最佳处理方式.流式细胞术检测结果显示凋亡细胞数量呈时间依赖性增加,并在48 h凋亡最为显著;凋亡相关蛋白PARP、caspase-3以及Cleaved-caspase-3均呈时间依赖性活化.以2 μg/mL桃儿七处理细胞48 h后,K562细胞形态发生不规则改变,出现核碎裂、溶解等凋亡特征;BCR/ABL、p-BCR/ABL、STAT5、p-STAT5表达显著降低.结论 藏药桃儿七能够促进慢性粒细胞白血病K562细胞凋亡,其机制可能通过抑制BCR/ABL-STAT5信号通路并激活线粒体凋亡途径来实现.

Effects of Sinopodophyllum hexundrum on apoptosis in K562 cells

Objective To investigate the effects of Sinopodophyllum hexundrum on apoptosis in K562 cells.Methods K562 cells were treated with Sinopodophyllum hexundrum at different concentrations and for different lengths of time to determine the optimal conditions of SinoPodophyllum hexandrum treatment for K562 cells using CCK8 assay.The cell apoptotic rate was detected by flow cytometry,and the cell morphology and nuclear morphology of K562 cells were observed with Wright staining and DPAI staining,respectively.The protein expressions of BCR/ABL,p-BCR/ABL,STAT5,p-STAT5 and the apoptosis-related proteins PARP,caspase-3 and cleaved-caspase-3 were determined with Western blotting.Results The ceil proliferation was inhibited in a concentration-and time-dependent manner by 1,2,and 3 μg/mL Sinopodophyllum hexundrum.The treatment was optimal with a Sinopodophyllum hexundrum concentration of 2 μg/mL a treatment time of 48 h,and the cell apoptotic rate increased in a time-dependent manner and significantly increased at 48 h (P＜0.001).The expression of apoptosis-related proteins PARP,caspase-3 and cleaved-caspase-3 were also activated in a time-dependent manner.The cells showed typical apoptotic changes after treatment with 2 μg/mL Sinopodophyllum hexundrum for 48 h with significantly reduced expressions of BCR/ABL,p-BCR/ABL,STAT5,AND p-STATS.Conclusion Sinopodophyllum hexundrum promotes K562 cell apoptosis possibly by inhibiting BCR/ABL-STAT5 survival signal pathways and activating the mitochondrion-associated apoptotic pathways.