Wortmannin抑制PI3K途径对bcr/abl基因介导的白血病细胞增殖和凋亡抗性影响的研究

A Study on the Efffects of Phosphatidylinositol-3 Kinase Inhibitor Wortmannin on Leukemia Cells Proliferation and Apoptosis Resistance Mediated by bcr/abl

目的探讨磷酰肌醇-3激酶(PI3K)途径在K562、NB4和HL60细胞增殖和凋亡抗性中的不同作用。方法用磷酰肌醇-3激酶(PI3K)特异抑制剂Wortmannin(WT)抑制PI3K活性,经细胞生长曲线测定、半固体集落形成实验、流式细胞膜联蛋白V(Annexin-V-Flous)标记技术检测细胞凋亡百分比和凋亡指数。观察K562、NB4和HL60细胞增殖能力及凋亡抗性的变化。结果K562、NB4和HL60细胞在24、48、72h的增殖抑制率分别为41.33%、57.46%、65.85%和26.29%、5.51%、2.10%及32.14%、17.14%、13.14%。生长曲线显示WT抑制PI3K途径可显著抑制K562细胞的增殖,对NB4和HL60细胞增殖无明显影响。K562、NB4和HL60细胞加和不加WT,培养14d后的集落形成率分别为16.15%和7.60%,5.90%和6.10%,6.60%和6.40%。集落形成抑制率为52.94%、3.39%和3.03%。K562、NB4和HL60细胞加WT、AraC、WT+AraC作用24h后的凋亡细胞百分比分别为(17.27±1.94)%、(23.25±14.12)%、(17.60±1.27)%;(17.00±3.36)%、(20.55±8.57)%、(22.07±5.62)%;(27.60±4.36)%、(17.56±9.28)%、(20.50±7.97)%。凋亡指数分别为(6.88±2.66)、(7.79±2.75)、(3.03±0.56);(8.91±3.86)、(9.35±4.19)、(3.79±0.93);(13.39±4.49)、(7.61±6.35)、(5.27±2.69)。结论WT可以通过抑制PI3K通?

Objective To investigate the effects of phosphatidylinositol-3 Kinase (PI3K) inhibitor wortmannin(WT) on leukemia cells proliferation and apoptosis resistance. Methods PI3K specific inhibitor WT was used to treat leukemia cell lines K562, NB4 and HL60. The changes of proliferation and apoptosis resistance in K562, NB4 and LH60 cells were observed. Results Proliferation curve and colony formation test showed that WT could obviously inhibit K562 cells proliferation and colony formation rate (P<0.05), but had not significant effects on the proliferation and clony formation rate of NB4 and LH60(P>0.05). FCM analysis using fluorescent labelled annexin-V antibody showed that WT could promote K562 cells apoptosis induced by AraC, but had not significant effects on AraC-induced cells apoptosis of NB4 and HL60. Conclusion WT can inhibit the proliferation and apoptosis resistance of K562. PI3K signaling pathway may be involved in leukemia cell proliferation and apoptosis resistance mediated by bcl/abl gene. [