全反式维甲酸联合小剂量阿糖胞苷对急性髓系白血病细胞磷脂酰肌醇3激酶/蛋白质丝氨酸苏氨酸激酶信号通路的作用机制研究OACSTPCD
Mechanism of all-trans retinoic acid combined with low-dose cytarabine on PI3K/AKT signaling pathway of acute myeloid leukemia cells
目的:探讨全反式维甲酸(ATRA)联合小剂量阿糖胞苷(Ara-C)对急性髓系白血病细胞磷脂酰肌醇3激酶(PI3K)/蛋白质丝氨酸苏氨酸激酶(AKT)信号通路的作用机制.方法:选取人急性髓系白血病细胞HL-60,分为空白对照组(未经任何处理的HL-60细胞)、Ara-C组(加入0.5 μmol/L Ara-C)、ATRA组(加入2 μmol/L ATRA)、ATRA+Ara-C 组(加入 2 μmol/L ATRA 和 0.5 μmol/L Ara-C),继续培养 24、48、72 h,采用 CCK8 检测 HL-60 细胞活力,AnnexinV 双染法检测 HL-60 细胞凋亡,qRT-PCR 检测 PI3K、AKT-mRNA 表达,Western blot 检测 PI3K/AKT信号通路相关蛋白表达,并进行细胞形态学观察.结果:空白对照组HL-60细胞活力高于Ara-C+ATRA组、Ara-C组、ATRA 组,Ara-C 组、ATRA 组 HL-60 细胞活力高于 Ara-C+ATRA 组(均 P＜0.05).Ara-C+ATRA 组HL-60细胞凋亡率高于Ara-C组、ATRA组,Ara-C组、ATRA组HL-60细胞凋亡率高于空白对照组(均P＜0.05).HL-60细胞胞体多呈圆形,可见瘤状突起,胞核多为类圆形,Ara-C组、ATRA组染色质凝聚,颜色变深,部分胞核变小,Ara-C+ATRA组染色质凝聚,颜色变深,可见核固缩、核碎裂.空白对照组HL-60细胞PI3K、AKT mRNA表达高于 Ara-C 组、ATRA 组,Ara-C 组、ATRA 组 HL-60 细胞 PI3K、AKT mRNA 表达高于 Ara-C+ATRA 组(均P＜0.05).空白对照组HL-60细胞P-PI3K、P-AKT蛋白表达高于Ara-C组、ATRA组,Ara-C组、ATRA组HL-60细胞P-PI3K、P-AKT蛋白表达高于Ara-C+ATRA组(均P＜0.05).结论:Ara-C+ATRA能通过抑制PI3K/AKT信号通路激活,促进HL-60细胞凋亡.
Objective:To investigate the mechanism of all-trans retinoic acid(ATRA)combined with low-dose cytarabine(Ara-C)on the signaling pathway of phosphatidylinositol 3 kinase(PI3K)/protein serine threonine kinase(AKT)in acute myeloid leukemia cells.Methods:Human acute myeloid leukemia cells HL-60 were selected.They were divided into blank control group(HL-60 cells without any treatment),Ara-C group(adding 0.5 μmol/L Ara-C),ATRA group(adding 2 μmol/L ATRA),ATRA+Ara-C group(adding 2 μmol/L ATRA and 0.5 μmol/L Ara-C).Culture continued for 24,48 and 72 hours.CCK8 was used to detect the viability of HL-60 cells,AnnexinV double staining was used to detect the apoptosis of HL-60 cells,qRT-PCR was used to detect the mRNA expression of PI3K and AKT,and Western blot was used to detect the protein expression of PI3K/AKT signaling pathway.The cell morphology was observed.Results:The activity of HL-60 cells in blank control group was higher than that in Ara-C+ATRA group,Ara-C group and ATRA group,and the activity of HL-60 cells in Ara-C and ATRA groups was higher than that in Ara-C+ATRA group(all P＜0.05).The apoptosis rate of HL-60 cells in Ara-C+ATRA group was higher than that in Ara-C and ATRA groups,and the apoptosis rate of HL-60 cells in Ara-C and ATRA groups was higher than that in blank control group(all P＜0.05).The body of HL-60 cells was mostly round with nodular pro-trusion,and the nuclei were mostly round like.The chromatin in the Ara-C and ATRA groups was condensed,the color became darker,and some of the nuclei became smaller.The chromatin in the Ara-C+ATRA group was con-densed,the color became darker,and the nuclear pyknosis and nuclear fragmentation were observed.The expressions of P13K and AKT mRNA in HL-60 cells of blank control group were higher than those in Ara-C and ATRA groups,and the expressions of PI3K and AKT mRNA in HL-60 cells of Ara-C and ATRA groups were higher than those in Ara-C+ATRA group(all P＜0.05).The expressions of P-PI3K and P-AKT in HL-60 cells of blank control group were higher than those in Ara-C and ATRA groups,and the expressions of P-PI3K and P-AKT in HL-60 cells of Ara-C and ATRA groups were higher than those in Ara-C+ATRA group(all P＜0.05).Conclusion:Ara-C+ATRA can promote the apoptosis of HL-60 cells by inhibiting the activation of PI3K/AKT signaling pathway.
杨白梅;鲁猛;骆思君;王志华;伍华英;王芳;赵耀顺
华北石油管理局总医院普通内科,河北任丘 062550廊坊市中医医院,河北廊坊 065800华北石油二部医院内科,河北任丘 062552华北石油中医医院内科,河北任丘 062550华北石油管理局总医院血液内科,河北任丘 062550
临床医学
全反式维甲酸阿糖胞苷急性髓系白血病HL-60细胞磷脂酰肌醇3激酶/蛋白质丝氨酸苏氨酸激酶信号通路细胞凋亡
All-trans retinoic acidCytarabineAcute myeloid leukemiaHL-60 cellsPI3K/AKT signaling pathwayApoptosis
《陕西医学杂志》 2024 (001)
28-31,36 / 5
河北省医学科学研究计划课题(20201252)
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