摘要
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.
作者
高晓艳
刘梅
田小清
吕润林
鲁英娟
GAO Xiao-yan;LIU Mei;TIAN Xiao-qing;LV Run-lin;LU Ying-juan(Department of Hematology,1.Yulin NO.2 Hospital,Yulin 719000;First Affiliated Hospital of Air Force Military Medical University,Xi'an,710032,Shaanxi,China)
出处
《川北医学院学报》
CAS
2019年第5期500-503,共4页
Journal of North Sichuan Medical College
基金
陕西省自然科学基金(ZRY201609026)
