RNAi干预急性髓系白血病对p38信号通路的影响  

Effect of RNAi on p38 pathway in acute myeloid leukemia

作　　者：王昭[1,2] 刘蕾 王柏欣[1] 王彬杉[1] 李晶 马淑霞[1] WANG Zhao;LIU Lei;WANG Baixin;WANG Binshan;LI Jing;MA Shuxia(College of Basic Medical Sciences,Jiamusi University,Jiamusi,Heilongjiang 154007,China)

机构地区：[1]佳木斯大学基础医学院,黑龙江佳木斯154007 [2]肇庆医学高等专科学校,广东肇庆526020

出　　处：《中国微生态学杂志》2019年第6期651-655,共5页Chinese Journal of Microecology

基　　金：黑龙江省教育厅骨干教师项目(1152G040)

摘　　要：目的探讨RNAi技术治疗急性髓系白血病(acute myeloid leukemia,AML)对p38信号通路的影响。方法以NC(HL-60细胞)和HK(含siRNA-FLT3的阴性对照的HL-60细胞)、F1(实验组,含siRNA-FLT3的HL-60细胞,本实验室已实验证明对FLT3有干扰作用)三种细胞株为研究对象,分为药物组(信号通道抑制作用组)和对照组(未加信号通道抑制剂)。通过RT-PCR和Western blot检测对照组和药物组p38 mRNA水平和蛋白表达的变化,MTT和FCM分别测定细胞活性和细胞凋亡率的变化。结果药物组、对照组组内比较:以NC做参照,F1能诱导p38 mRNA水平和蛋白表达下降、从而抑制细胞的增殖,促进细胞凋亡,差异有统计学意义(均P<0.05);HK与NC之间差异无统计学意义(P>0.05);与对照组相比,药物组p38 mRNA水平和蛋白表达下降,细胞活性下降,细胞凋亡率增加,差异有统计学意义(均P<0.05);药物组的F1与药物组的NC、药物组的F1与对照组的F1分别比较,差异有统计学意义(均P<0.05)。结论 p38信号通路是RNAi技术干扰HL-60细胞FLT3基因表达引起细胞凋亡的途径之一,SB203580(p38 MAPK抑制剂)对其有协同增效作用。Objective To investigate the effect of siRNA-FLT3 on p38 signaling pathway in acute myeloid leukemia(AML). Methods The cell lines of NC(HL-60 cells),HK(HL-60 cells containing the negative control of siRNA-FLT3)and F1(experiment group,HL-60 cells containing the siRNA-FLT3 proved to interfere the expression of FLT3 very well in our laboratory)were used as the subjects and divided into the drug group(signaling pathway inhibition group)and control group(with no signaling pathway inhibitor).RT-PCR and Western blot were used to detect the changes of p38 mRNA level and protein expression.MTT and FCM were used to determine the changes of cell viability and percentage of apoptosis. Results Compared with NC,the expression of p38 mRNA and protein declined in both drug group and control group,which inhibited cell proliferation and promoted cell apoptosis.The differences were statistically significant(all P<0.05).There were no statistical differences between HK and NC(all P>0.05).Compared with the control group,the p38 mRNA level and protein expression decreased in the drug group;the cell viability declined while the apoptosis increased.The differences were statistically significant(all P<0.05).The differences between F1 and NC in drug group were statistically significant(all P<0.05),so were those between F1 in drug group and F1 in control group. Conclusion The interference of FLT3 gene through p38 signaling pathway can cause apoptosis of HL-60.SB203580(p38 MAPK inhibitor)has a synergistic effect.

关 键 词：RNA干扰 FLT3 P38 急性髓系白血病 

分 类 号：R733.7[医药卫生—肿瘤]