微小RNA-150靶向c-Myb表达对T淋巴细胞白血病Jurkat细胞增殖的影响  

MiR-150 targeting c-Myb expression inhibits the proliferation of Jurkat cells

作　　者：杨莉[1] 刘炜[1] 李彦格 管玉洁[1] 毛彦娜[1] YANG Li;LIU Wei;LI Yange;GUAN Yujie;MAO Yanna(Department of Hematological Oncology,Zhengzhou Children's Hospital,Henan Children's Hospital,Children's Hospital Affiliated to Zhengzhou University,Zhengzhou,Henan 450000,China)

机构地区：[1]郑州儿童医院、河南省儿童医院、郑州大学附属儿童医院血液肿瘤科,河南郑州450000

出　　处：《安徽医药》2022年第8期1615-1618,共4页Anhui Medical and Pharmaceutical Journal

基　　金：2018年度河南省医学科技攻关计划(2018020679)。

摘　　要：目的探讨微小RNA-150(miR-150)靶向c-Myb表达对人T淋巴细胞白血病(T-LL)Jurkat细胞增殖的影响。方法体外培养人Jurkat细胞,分为空白对照组(不转染)、阴性对照组(转染negative control-miR-150)和miR-150过表达组(转染hsamiR-150 mimic)。实时荧光定量PCR(qRT-PCR)法检测Jurkat细胞中miR-150及c-Myb表达;人胆囊收缩素/缩胆囊素八肽(CCK-8)法检测Jurkat细胞增殖;流式细胞仪检测各组Jurkat细胞凋亡情况;双荧光素酶报告基因实验检测miR-150和c-Myb的靶向关系;蛋白质印迹法(Western blotting)检测各组Jurkat细胞中c-Myb、增殖细胞核抗原(PCNA)、Bcl-2相关X蛋白(Bax)蛋白表达情况。结果空白对照组与阴性对照组Jurkat细胞中miR-150表达水平、c-Myb mRNA及蛋白表达水平、OD值、细胞凋亡率、及PCNA、Bax蛋白水平差异无统计学意义(P>0.05);与空白对照组相比,miR-150过表达组Jurkat细胞中miR-150表达水平[(1.42±0.14)比(1.03±0.09)]、凋亡率[(20.79±1.15)%比(8.76±0.74)%]、Bax蛋白表达水平[(0.65±0.18)比(0.42±0.13)]显著升高(P<0.05),c-Myb mRNA[(0.66±0.14)比(0.98±0.09)]及蛋白表达水平[(0.37±0.06)比(0.64±0.12)]、OD值[(0.25±0.06)比(0.46±0.09)]、PCNA蛋白表达水平[(0.33±0.07)比(0.56±0.11)]显著降低(P<0.05)。双荧光素酶报告基因实验显示,c-Myb是miR-150的靶基因。结论miR-150靶向c-Myb表达抑制Jurkat细胞增殖并诱导其凋亡。Objective To investigate the effect of microRNA-150(miR-150)targeting c-Myb expression on the proliferation of Jurkat cells.Methods Human Jurkat cells were cultured in vitro and assigned into blank control group(no transfection),negative control group(transfected with negative control-miR-150)and miR-150 overexpression group(transfected with hsa-miR-150 mimic).The expressions of miR-150 and c-Myb in Jurkat cells were detected by real-time fluorescent quantitative PCR(qRT-PCR);the proliferation of Jurkat cells was detected by cholecystokinin octapeptide(CCK-8);the apoptosis of Jurkat cells was detected by flow cytometry;the targeting relationship between miR-150 and c-Myb was detected by double luciferase reporter gene assay;and Western blotting(WB)was used to detect the protein expressions of c-Myb,proliferating cell nuclear antigen(PCNA)and Bcl-2-associated X-protein Bax in Jurkat cells.Results There were no significant differences in the expression levels of miR-150,c-Myb mRNA and protein,OD value,apoptosis rate,protein expression levels of PCNA and Bax in Jurkat cells between blank control group and negative control group(P>0.05).Compared with blank control group,the expression level of miR-150[(1.42±0.14)vs.(1.03±0.09)],apoptosis rate[(20.79±1.15)%vs.(8.76±0.74)%]and Bax protein[(0.65±0.18)vs.(0.42±0.13)]in Jurkat cells of miR-150 overexpression group were significantly higher(P<0.05),while the expression levels of c-Myb mRNA[(0.66±0.14)vs.(0.98±0.09)]and protein[(0.37±0.06)vs.(0.64±0.12)],OD value[(0.25±0.06)vs.(0.46±0.09)]and protein expression level of PCNA[(0.33±0.07)vs.(0.56±0.11)]were significantly lower(P<0.05).Double luciferase reporter gene experiment results showed that c-Myb was the target gene of miR-150.Conclusion The miR-150 targeting c-Myb expression inhibits the proliferation and induces the apoptosis of Jurkat cells.

关 键 词：白血病 T细胞 微小RNA-150 C-MYB JURKAT细胞 细胞增殖 

分 类 号：R733.7[医药卫生—肿瘤]