摘要
目的观察磷脂酰肌醇3-激酶/丝氨酸苏氨酸激酶(PI3K/Akt)特异性抑制剂LY294002对人套细胞淋巴瘤Jeko-1细胞株细胞增殖、凋亡及Notch1信号通路的影响。方法四甲基偶氮唑盐(MTT)法检测细胞增殖抑制率;流式细胞仪检测细胞凋亡变化;Western blot检测凋亡相关蛋白Bcl-2、Bax、Caspase-3、Caspase-9及信号通路相关蛋白pAkt、Notch1、HES1的变化。结果 LY294002能明显抑制Jeko-1细胞的增殖;经0、5、10和20μmol/L的LY294002作用24h后,Jeko-1细胞的凋亡率分别为(3.25±1.27)%、(11.34±2.35)%、(22.81±2.74)%、(43.61±3.48)%,差异有统计学意义(P<0.01);Western blot检测发现凋亡相关蛋白Bcl-2表达下降,Bax、Caspase-3、Caspase-8、Caspase-9表达上升,PI3K/Akt信号通路相关蛋白p-Akt磷酸化水平下降,Notch1信号通路相关蛋白Notch1、HES1表达下降。结论LY294002可以特异性抑制PI3K/Akt信号通路,也可下调Notch1信号通路的活性,抑制Jeko-1细胞增殖,促进细胞凋亡。
Objective To investigate the effects of the specific inhibitor of phosphatidylinositol 3-kinase/Akt（PI3K/Akt）,LY294002,on the proliferation and apoptosis of Jeko-1cell line of mantle cell lymphoma（MCL）,and on the expressions of Notch1 signaling pathway-related proteins.Methods Jeko-1cells were cultured with different concentrations of LY294002.Cell growth was determined by MTT.Cell apoptosis was analyzed by flow cytometry.The expressions of apoptosis-related proteins（Bcl-2,Bax,Caspase-3and Caspase-9）and Notch1 signaling pathway-related proteins（p-Akt,Notch1 and HES1）were detected by Western blot.Results LY294002 could inhibit the proliferation of Jeko-1cells in a time-and dose-dependent manner.The cell apoptotic rates of Jeko-1cells treated with 0,5,10 and 20μmol/L of LY294002 for 24h were（3.25±1.27）%,（11.34±2.35）%,（22.81±2.74）% and（43.61±3.48）%,respectively,and there were significant differences（P0.01）.Western blot showed that the expression of Bcl-2 was down-regulated,those of Bax,Caspase-3,Caspase-8and Caspase-9up-regulated,the phosphorylation level of the PI3K/Akt signaling pathway-related protein p-Akt decreased and the expressions of Notch1 signaling pathway-related proteins Notch1 and HES1reduced.Conclusion LY294002 can suppress the growth and induce the apoptosis of Jeko-1cells by specifically inhibiting the PI3K/Akt signaling pathway and down-regulating the activity of Notch1 signaling pathway.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2015年第2期156-159,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
福建省引进重大研发机构资助项目(No.2012I2004)
