摘要
目的研究微小RNA-21(mi R-21)靶向Fas配体(Fas L)调控急性髓系白血病细胞增殖、凋亡的作用。方法培养急性髓系白血病细胞THP-1,转染阴性对照(NC)模拟物、mi R-21模拟物、NC抑制物、mi R-21抑制物后采用荧光定量PCR法检测mi R-21及Fas L、含半胱氨酸天冬氨酸蛋白水解酶-8(Caspase-8)、Caspase-3的表达量,采用四氮唑蓝盐化合物(MTS)法检测细胞增殖活力的OD540值,采用末端脱氧核苷酸转移酶介导的d UTP缺口末端标记测定法(TUNEL)法检测细胞凋亡率,采用荧光素酶报告基因验证mi R-21与Fas L基因m RNA 3’UTR的靶向结合。结果 mi R-21模拟物组细胞中mi R-21的表达量及OD540值均明显高于NC模拟物组,细胞中Fas L、Caspase-8、Caspase-3的m RNA表达量、凋亡率、Fas L基因m RNA 3’UTR荧光素酶报告基因的荧光活性明显低于NC模拟物组,差异均有统计学意义(P<0.05);mi R-21抑制物组细胞中mi R-21的表达量及OD540值均明显低于NC抑制物组,细胞中Fas L、Caspase-8、Caspase-3的m RNA表达量及凋亡率明显高于NC抑制物组,差异均有统计学意义(P<0.05)。结论 miR-21能够促进急性髓系白血病细胞增殖、抑制细胞凋亡,且该作用与靶向抑制FasL有关。
Objective To study the effects of miR-21 on regulating proliferation and apoptosis of acute myeloid leukemia cells via targeting FasL.Methods THP-1 cells were cultured and transfected with negative control(NC) mimic,miR-21 mimic,NC inhibitor and miR-21 inhibitor.The expressions levels of miR-21,FasL,Caspase-8 and Caspase-3 were detected by fluorescence quantitative PCR.The OD540 value of cell proliferation activity was detected by MTS method,and the apoptotic rate was detected by TUNEL method.Luciferase reporter gene was used to validate the targeted binding of miR-21 to FasL gene 3’UTR.Results The expression of miR-21 and the OD540 value of miR-21 mimic group were significantly higher than those in NC mimic group,the mRNA expression of FasL,Caspase-8,Caspase-3,apoptotic rate and the fluorescence activity of luciferase reporter gene containing FasL gene 3’UTR of mimic group were significantly lower than those in NC mimic group(P<0.05).The expression of miR-21 and the OD540 value of miR-21 inhibitor group were significantly lower than those in NC inhibitor group,the mRNA expression of Fast,Caspase-8,Caspase-3 and the apoptotic rate were significantly higher than those in NC inhibitor group(P<0.05).Conclusion miR-21 could promote the proliferation and inhibit apoptosis of acute myeloid leukemia cells,and this effect was related to targeted inhibition of FasL.
作者
郑研
高秋英
牛奔
张维华
李岚
ZHENG Yan;GAO Qiu-ying;NIU Ben;ZHANG Wei-hua;LI Lan(Department of Hematology,People's Hospital of West China,Xi'an,Shaanxi 710068,China)
出处
《热带医学杂志》
CAS
2020年第10期1300-1303,共4页
Journal of Tropical Medicine
