miR-27a-3p调控XIAP表达对急性T淋巴细胞白血病Jurkat细胞增殖和凋亡的影响  

Effects of miR-27a-3p regulating XIAP expression on proliferation and apoptosis of Jurkat cells in acute T lymphocytic leukemia

作　　者：刘牧 杨夏茵 薄海美 LIU Mu;YANG Xiayin;BO Haimei(Department of Pediatrics,Affiliated Hospital of North China University of Science and Technology,Tangshan,Hebei 063000,China;Department of Cardiovascular Disease,School of Clinical Medicine,North China University of Science and Technology,Tangshan,Hebei 063000,China)

机构地区：[1]华北理工大学附属医院儿科,河北唐山063000 [2]华北理工大学临床医学院心血管,河北唐山063000

出　　处：《重庆医学》2021年第9期1446-1450,共5页Chongqing medicine

基　　金：河北省政府资助专科能力建设和专科带头人培养项目(361036)。

摘　　要：目的探讨微小RNA(miR)-27a-3p对急性T淋巴细胞白血病(T-ALL)Jurkat细胞增殖和凋亡的影响及其机制。方法将体外培养的T-ALL Jurkat细胞分为未转染组(未处理)、miR-NC组(转染阴性对照miR-NC)和miR-27a-3p组(转染miR-27a-3p模拟物),采用实时荧光定量PCR(qRT-PCR)检测细胞中miR-27a-3p表达水平,噻唑蓝(MTT)法检测细胞增殖,流式细胞仪检测细胞凋亡,Western blot检测细胞中B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和X染色体连锁的凋亡抑制基因(XIAP)蛋白表达水平,比色法检测细胞半胱氨酸天冬氨酸蛋白酶(caspase)-3活性,双荧光素酶报告基因(DLR)实验检测miR-27a-3p和XIAP的靶向关系。结果与未转染组或miR-NC组比较,miR-27a-3p组细胞中miR-27a-3p表达水平、细胞凋亡率、细胞中Bax表达水平和caspase-3活性均明显升高,而细胞增殖活力和细胞中Bcl-2、XIAP蛋白表达水平均明显降低(P<0.05);miR-NC组和未转染组上述各指标比较差异均无统计学意义(P>0.05)。DLR实验证实miR-27a-3p可与XIAP靶向结合。结论miR-27a-3p可抑制T-ALL Jurkat细胞增殖并诱导细胞凋亡,其作用机制可能与靶向调控XIAP表达有关。Objective To investigate the effects of microRNA(miR)-27a-3p on the proliferation and apoptosis of Jurkat cells of acute T lymphocytic leukemia(T-ALL)and its mechanism.Methods The in vitro cultured Jurkat cells of T-ALL were divided into the untransfected group(without treated),miR-NC group(transfected with negative control miR-NC)and miR-27a-3p group(transfected with miR-27a-3p analog).The expression level of miR-27a-3p in cells was detected by real-time fluorescence quantitative PCR,the cell proliferation was detected by MTT,and the apoptosis was detected by flow cytometry,Western blot was used to detect the protein expression levels of B-lymphoma-2(Bcl-2),Bcl-2 related X protein(Bax)and X-linked inhibitor of apoptosis protein gene(XIAP),the activity of cysteinyl aspartate specific proteinase(caspase-3)was detected by colorimetry,and the target relationship between miR-27a-3p and XIAP was detected by double luciferase reporter gene assay.Results The expression level of miR-27a-3p,apoptosis rate,Bax protein expression level and caspase-3 activity in the miR-27a-3p group were significantly increased compared with those in the non transfected group or miR-NC group,however,the cellular proliferation activity and the expression levels of Bcl-2 and XIAP proteins in the cells were significantly decreased(P<0.05);and there was no statistically significant difference in the above indexes between the miR-NC group and untransfected group(P>0.05).In addition,the double luciferase reporter(DLR)experiment confirmed that miR-27a-3p could have the targeted combination with XIAP.Conclusion MiR-27a-3p can inhibit the proliferation of T-ALL Jurkat cells and induce the apoptosis,and its mechanism may be related to the targeted regulation of XIAP expression.

关 键 词：急性T淋巴细胞白血病 微小RNA-27a-3p 细胞增殖 细胞凋亡 X染色体连锁的凋亡抑制基因 

分 类 号：R733.7[医药卫生—肿瘤]