Wnt信号通路抑制对急性白血病天门冬酰胺酶耐药性的影响

Effects of Wnt signaling pathway inhibition on asparaginase resistance in acute leukemia

目的:探讨Wnt信号通路抑制对急性淋巴细胞白血病(ALL)天门冬酰胺酶(Asp)耐药性的影响。方法:人ALL细胞株THP-1分为对照组、Asp组与si-Wnt组,Asp组与si-Wnt组分别转染siRNA阴性对照与Wnt siRNA,转染后48 h,两组均加入Asp(终浓度为0.25μM),对照组只加生理盐水。MTT法检测细胞增殖,流式细胞仪检测细胞凋亡与细胞周期,Western-blot法检测蛋白表达水平。结果:Asp加药后24 h、48 h、72 h,si-Wnt组细胞增殖抑制率和细胞凋亡率均高于对照组和Asp组,且Asp组高于对照组(P<0.05)。Asp加药后72 h,si-Wnt组的G0/G1期细胞比例、Wnt及AKT蛋白表达均低于对照组与Asp组,Asp组低于对照组(P<0.05)。而si-Wnt组G2/M期细胞比例高于对照组与Asp组,且Asp组高于对照组(P<0.05)。结论:Wnt信号通路抑制可降低ALL细胞对Asp的耐药性,抑制AKT蛋白的表达,阻滞细胞周期,促进ALL细胞的凋亡,抑制其增殖。

Objective:To investigate the effects of Wnt signaling pathway inhibition on asparaginase(Asp)resistance in acute lymphoblastic leukemia(ALL).Methods:Human ALL cell line THP-1 were divided into three groups:control group,Asp group and si-Wnt group.Asp group and si-Wnt group were transfected with siRNA negative control and Wnt siRNA,respectively.The Asp(final concentration 0.25μM)were added 48 h after transfection in Asp group and si-Wnt group.The control group only added normal saline.Cell proliferation were detected by MTT assay,apoptosis and cell cycle were detected by flow cytometry,and protein expression were detected by Western-blot method.Results:At 24h,48h and 72h after Asp administration,the cell proliferation inhibition rate and apoptosis rate of si-Wnt group were significantly higher than those of control group and Asp group,and Asp group was higher than control group(P<0.05).At 72 h after Asp administration,the proportion of G0/G1 phase cells,expressions of Wnt and AKT proteins in si-Wnt group were significantly lower than those in control group and Asp group,and Asp group was lower than control group(P<0.05).The proportion of G2/M phase cells in si-Wnt group was significantly higher than that in control group and Asp group,and Asp group was higher than control group(P<0.05).Conclusion:Inhibition of Wnt signaling pathway can reduce the resistance of ALL cells to Asp,inhibit the expression of AKT protein,arrest the cell cycle,promote the apoptosis of ALL cells,and inhibit its proliferation.