磷脂酰肌醇3-激酶抑制剂对分化的HL-60细胞凋亡的诱导

Apoptosis of differentiated HL-60 cells induced by inhibitor of phosphatidylinositol 3-kinase

目的 探讨磷脂酰肌醇 3 -激酶 (phosphatidylinositol 3 -kinase ,PI3 -K)抑制剂对分化的人急性粒细胞白血病HL -60细胞生存的影响。方法 流式细胞仪检测分化抗原CD11b的表达 ,以确定分化程度 ;DNA梯形条带电泳确定凋亡 ;流式细胞仪分选分化与未分化细胞 ,并检测凋亡。结果 1μmol/L全反式维甲酸 (all-transretinoicacid ,ATRA)ATRA处理HL -60细胞 12 ,2 4,48h后 ,CD11b的阳性率分别为 2 0 2 % ,3 1 1% ,64 5 % ;经ATRA处理过的HL -60细胞中加入PI3 -K的抑制剂wortmannin( 1μmol/L) 2h ,在琼脂糖凝胶电泳上可见凋亡细胞特征性的“梯形”带 ;流式细胞仪分选出分化与未分化细胞后 ,在分化细胞中加入wortmannin( 1μmol/L) 2h ,PI染色流式细胞仪直方图上可见亚二倍体峰。结论 全反式维甲酸诱导分化的HL -60细胞的生存是依赖于PI3

Objective To study the effect of wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3-K), on the survival of differentiated human acute promyloid HL-60 cells. Methods\ Flow cytometry was used to detect cell surface expression of CD11b; Apoptosis was detected by DNA electrophoresis; Flow cytometry was also used to sort differentiated and undifferentiated HL-60 cells and to detect apoptotic cells. Results\ The proportion of CD11b-expressing HL-60 cells cultured with ATRA(all-trans retinoic acid) for 12 h, 24 h and 48 h is 20 2%,31 1% and 64 5%, respectively; Internuleosomal DNA damage and DNA ladder in HL-60 cells treated by ATRA was detected 2 h after addition of wortmannin, but not in non-ATRA-treated cells; Cells were seperated and selected into differentiated and non-differentiated cells by flow cytometry, and differentiated ones exhibited a sharp subG 1 peak compared to undifferentiated cells following wortmannin treatment. The apoptotic indexes were 62 6%,66 5% and 89 8% respectively. Conclusion\ The survival of differentiated HL-60 cells induced by ATRA depends on the PI3-K pathway to transduce survival signal.