SHIP1调控STAT3信号通路对白血病Jurkat细胞活力和凋亡的影响

摘要

目的:探讨含SH2结构域的肌醇5-磷酸酶1(SHIP1)通过调控信号转导及转录激活因子3(STAT3)信号通路对人白血病细胞活力和凋亡的影响.方法:以白血病Jurkat细胞为研究对象,将转染空载体和SHIP1过表达载体的细胞分别记为阴性对照(NC)组和SHIP1组,同时以不作处理的细胞为空白对照(control)组,用real-time PCR和Western blot检测转染效果,MTT法检测细胞活力,流式细胞术检测细胞凋亡,Western blot检测细胞中活化的caspase-3(cleaved caspase-3)、STAT3和磷酸化STAT3(p-STAT3)的蛋白水平.同时,用STAT3信号通路抑制剂AG490作用于control组和SHIP1组细胞,分别记为control+AG490组和SHIP1+AG490组,再观测上述指标的变化.结果:分别与control组和NC组比较,SHIP1组SHIP1的mRNA表达水平和蛋白水平均显著升高(P<0.05);细胞存活率显著降低(P<0.05);细胞凋亡率显著升高(P<0.05);cleaved caspase-3蛋白水平显著升高(P<0.05);p-STAT3蛋白水平显著降低(P<0.05).分别与control组和control+AG490组比较,SHIP1+AG490组的细胞存活率显著降低(P<0.05);cleaved caspase-3蛋白水平显著升高(P<0.05);p-STAT3蛋白水平显著降低(P<0.05);细胞凋亡率显著升高(P<0.05).结论:SHIP1能够通过抑制STAT3信号通路抑制人白血病细胞生长,促进白血病细胞凋亡.%AIM:To investigate the effect of SH2 domain-containing inositol 5-phosphatase 1(SHIP1)on the viability and apoptosis of leukemia cells by regulating signal transducer and activator of transcription 3(STAT3)signaling pathway.METHODS:Human leukemia Jurkat cells were transfected with null vector and SHIP1 overexpression vector in negative control(NC)group and SHIP1 group,respectively.The cells without treatment served as control group.The transfection efficiency was detected by real-time PCR and Western blot.The changes of the cell activity were measured by MTT assay.The apoptosis was detected by flow cytometry.The protein levels of cleaved caspase-3,STAT3 and p-STAT3 were determined by Western blot.The STAT3 signaling pathway inhibitor AG 490 was applied to the cells in control group and SHIP1 group as control +AG490 group and SHIP1+AG490 group,respectively,and the above indexes were also ana-lyzed.RESULTS:Compared with the control group and NC group ,the mRNA and protein expression levels of SHIP 1 in SHIP1 group were both upregulated ,the cell viability was increased ,the apoptotic rate was decreased ,the protein level of cleaved caspase-3 was upregulated ,and the protein level of p-STAT3 was decreased(P<0.05 ).Compared with control group and control +AG490 group,the cell viability in SHIP1+AG490 group was decreased ,the protein level of cleaved caspase-3 was increased the protein level of p-STAT3 was decreased ,and the apoptotic rate was increased(P<0.05 ). CONCLUSION:SHIP1 inhibits the viability and promotes the apoptosis of human leukemia Jurkat cells by inhibiting the STAT3 signaling pathway.