miRNA-126-3p调控人单核细胞白血病THP-1细胞的增殖和周期变化  

miRNA-126-3p regulates the proliferation and cycle change of human monocytic leukemia THP-1 cells

作　　者：遆刚 李莉莉[2] 李峰[3] Ti Gang;Li Lili;Li Feng(Department of Medical Record,Shanxi Bethune Hospital,Shanxi Academy of Medical Sciences,Tongji Shanxi Hospital,Taiyuan 030032,China;Department of Radiotherapy for Abdominopelvic tumors,Shanxi Province Cancer Hospital,Shanxi Hospital Affiliated to Cancer Hospital,Chinese Academy of Medical Sciences,Cancer Hospital Affiliated to Shanxi Medical University,Taiyuan 030013,China;Central Laboratory,Shanxi Province Cancer Hospital,Shanxi Hospital Affiliated to Cancer Hospital,Chinese Academy of Medical Sciences,Cancer Hospital Affiliated to Shanxi Medical University,Taiyuan 030013,China)

机构地区：[1]山西医学科学院、山西白求恩医院、同济山西医院病案室,太原030032 [2]山西省肿瘤医院、中国医学科学院肿瘤医院山西医院、山西医科大学附属肿瘤医院放腹盆科,太原030013 [3]山西省肿瘤医院、中国医学科学院肿瘤医院山西医院、山西医科大学附属肿瘤医院中心实验室,太原030013

出　　处：《白血病．淋巴瘤》2023年第12期740-744,共5页Journal of Leukemia & Lymphoma

基　　金：山西省科技厅基础研究计划(202203021221281);山西省卫健委"四个一批"重点攻关专项(2022XM28);山西省肿瘤医院科研创新团队建设项目(202003)。

摘　　要：目的探讨miRNA-126-3p(miR-126-3p)对人单核细胞白血病THP-1细胞增殖和周期的影响。方法采用实时荧光定量聚合酶链反应(PCR)检测miR-126-3p在THP-1细胞和从2022年5月至2023年1月山西省肿瘤医院收集的4名健康体检者外周血分离的单核细胞中的表达情况。构建过表达miR-126-3p载体,转染THP-1细胞(过表达miR-126-3p组),以转染空载体的THP-1细胞为对照。采用CCK-8法检测各组细胞增殖能力,采用流式细胞术检测细胞周期。应用TargetScan软件预测miR-126-3p靶基因为RGS3,通过双荧光素酶报告基因实验进行验证。采用蛋白质印迹法检测过表达miR-126-3p组THP-1细胞RGS3蛋白表达水平。结果PCR检测结果显示,与健康人外周血分离的单核细胞相比,THP-1细胞miR-126-3p表达水平低(P<0.05)。与对照组比较,过表达miR-126-3p组THP-1细胞在培养48 h和72 h后增殖能力均低(均P<0.05),且细胞G1期阻滞[G1期细胞比例:(58.2±2.8)%比(44.1±2.4)%,P<0.05]。双荧光素酶报告基因实验结果显示miR-126-3p与RGS3的3’’UTR结合。蛋白质印迹法检测显示,过表达miR-126-3p组THP-1细胞RGS3蛋白表达水平低于对照组。结论miR-126-3p可能通过抑制RGS3表达而抑制人单核细胞白血病THP-1细胞增殖和促进细胞G1期阻滞。Objective To investigate the effects of miRNA-126-3p(miR-126-3p)on proliferation and cycle of human monocytic leukemia THP-1 cells.Methods Real-time fluorescence quantitative polymerase chain reaction(PCR)was used to verify the expression of miR-126-3p in THP-1 cells and monocytes isolated from the peripheral blood of healthy check-ups collected from May 2022 to January 2023 in Shanxi Cancer Hospital.The miR-126-3p overexpression vector was constructed and transfected inyo THP-1 cells(miR-126-3p overexpression group),and THP-1 cells transfected with the empty vector were used as the control.The proliferative ability of cells in each group was detected by CCK-8 assay,and the cell cycle was detected by flow cytometry.TargetScan software was applied to predict the miR-126-3p target gene as RGS3,which was verified by dual luciferase reporter gene assay.The protein expression level of RGS3 in THP-1 cells of the miR-126-3p overexpression group was detected by Western blotting.Results The results of PCR showed that the expression level of miR-126-3p in THP-1 cells was low compared with that in monocytes isolated from peripheral blood of healthy individuals(P<0.05).Compared with the control group,THP-1 cells in the miR-126-3p overexpression group had low proliferative capacity after 48 h and 72 h of culture(both P<0.05),and the cells were blocked in G1 phase[proportion of cells in G1 phase:(58.2±2.8)%vs.(44.1±2.4)%,P<0.05].The results of dual luciferase reporter gene assay showed that miR-126-3p bound to the 3'UTR of RGS3.Western blotting assay showed that the RGS3 protein expression level in THP-1 cells of the miR-126-3p overexpression group was lower than that in the control group.Conclusions miR-126-3p may inhibit proliferation of human monocytic leukemia THP-1 cells and promote G1-phase blockade by inhibiting the expression of RGS3.

关 键 词：白血病 单核细胞 急性 miRNA-126-3p RGS3蛋白 人 细胞增殖 细胞周期 

分 类 号：R733.7[医药卫生—肿瘤]