LncRNA SNHG7通过抑制miR-186增强人急性髓系白血病细胞耐药性  

LncRNA SNHG7 enhances the drug resistance of human acute myeloid leukemia cells through inhibiting miR-186

作　　者：朱娟[1] 周英[2] 李颖佳[1] ZHU Juan;ZHOU Ying;LI Ying-jia(Department of Laboratory Medicine,the Third Xiangya Hospital of Central South University,Changsha 410013;Department of Clinical Laboratory,Changsha Central Hospital,Changsha 410004,China)

机构地区：[1]中南大学湘雅三医院检验科,湖南长沙410013 [2]长沙市中心医院检验科,湖南长沙410004

出　　处：《基础医学与临床》2021年第2期171-177,共7页Basic and Clinical Medicine

基　　金：湖南省卫生健康委科研计划(20201453);湖南省自然科学基金青年基金(2020JJ5877)。

摘　　要：目的探讨长链非编码RNA(lncRNA)SNHG7通过抑制miR-186对人急性髓系白血病(AML)细胞化疗耐药性的影响。方法取AML初诊患者、复发/难治患者及健康对照者的外周血;AML阿霉素敏感细胞(HL60)和AML阿霉素耐药细胞(HL60/ADM),采用荧光定量PCR检测外周血及细胞样本中SNHG7和miR-186的表达。CCK8法实验检测HL60和HL60/ADM细胞对阿霉素的敏感性并计算半数抑制浓度(IC 50)。在HL60/ADM细胞中转染sh-SNHG7、miR-186 mimic或miR-186 inhibitor,荧光定量PCR检测SNHG7和miR-186的表达,CCK8检测HL60/ADM对阿霉素的剂量反应和IC 50值。双荧光素酶报告基因实验检测SNHG7与miR-186的靶向结合。结果AML初诊者和复发/难治者血清中SNHG7的表达水平显著高于对照组,miR-186的趋势与SNHG7相反(P<0.01)。用不同浓度阿霉素处理后,HL60/ADM细胞活力显著高于HL60细胞(P<0.05),且阿霉素对HL60/ADM细胞的IC 50值3.36±0.65显著高于HL60细胞0.43±0.16(P<0.001)。HL60/ADM细胞中SNHG7显著高表达(P<0.05),miR-186显著低表达(P<0.05)。sh-SNHG7或miR-186 mimic转染HL60/ADM细胞后显著增加对阿霉素的敏感性,使IC 50值分别从3.17±0.61减少到2.30±0.31,3.22±0.62减少到2.16±0.33(P<0.05)。双荧光素酶报告基因实验证实SNHG7的吸附靶基因为miR-186。结论SNHG7通过抑制miRNA-186增强AML细胞化疗耐药性。Objective To investigate the effect of long non-coding RNA(lncRNA)SNHG7 on cell drug resistance through inhibiting microRNA-186(miR-186).Methods Peripheral blood was taken from newly diagnosed AML patients,recurrent/refractory AML patients and healthy controls.AML adriamycin sensitive cells(HL60)and AML adriamycin resistant cells(HL60/ADM)were used to detect the expression of SNHG7 and miR-186 in peripheral blood and cell samples by fluorescence quantitative PCR.The sensitivity of HL60 and HL60/ADM cells to adriamycin was determined by CCK8 assay and the 50% inhibitory concentration(IC 50)was calculated.Sh-SNHG7 or miR-186 mimic or miR-186 inhibitor was transfected into HL60/ADM cells.The expres-sion of SNHG7 and miR-186 was detected by fluorescence quantitative PCR,the dose response and IC 50 of HL60/ADM to adriamycin were detected by CCK8.Double luciferase reporter gene assay was used to detect the targeted binding of SNHG7 to miR-186.Results The expression of SNHG7 in the serum of newly diagnosed AML and recurrent/refractory AML patients was significantly higher than that of normal controls,and the trend of miR-186 was contrary to that of SNHG7(P<0.01).Compared with HL60 cells,HL60/ADM cells were treated with adriamycin at different concentrations,and the IC 50 value of adriamycin on HL60/ADM(3.36±0.65)cells was significantly higher than that of HL60 cells(0.43±0.16)(P<0.001).SNHG7 expression was significantly higher(P<0.05)in HL60/ADM cells and miR-186 was significantly lower(P<0.05).Transfection with sh-SNHG7 or miR-186 mimic in HL60/ADM cells increased the sensitivity to adriamycin and decreased the IC 50 value from 3.17±0.61 to 2.30±0.31 and 3.22±0.62 to 2.16±0.33,respectively(P<0.05).Dual-luciferase reporter assay confirmed that SNHG7 could bind miR-186.Conclusions SNHG7 enhances drug resistance of AML cells during chemotherapy through inhibition of miR-186.

关 键 词：SNHG7 miR-186 增殖 耐药 急性髓性白血病 

分 类 号：R733.71[医药卫生—肿瘤]