LncRNA MEG3通过下调miR-191影响表阿霉素对T细胞淋巴瘤细胞的增殖抑制和凋亡诱导作用  

The effect of LncRNA MEG3 on epirubicin-induced apoptosis and anti-proliferation of T-cell lymphoma cell by down-regulating miR-191

作　　者：聂成军 陈康银 陈仁利 陈琦 赖晓兰 唐宝佳 叶作东 NIE Cheng-jun;CHEN Kang-yin;CHEN Ren-li;CHEN Qi;LAI Xiao-lan;TANG Bao-jia;YE Zuo-dong(Department of Hematology,Rheumatism and Immunology,Ningde Municipal Hospital Affiliated to Ningde Normal University,Ningde Fujian 352100,China)

机构地区：[1]宁德师范学院附属宁德市医院血液风湿免疫科,福建宁德352100

出　　处：《毒理学杂志》2020年第6期466-471,共6页Journal of Toxicology

摘　　要：目的探讨长链非编码RNA(LncRNA)人母系表达基因3(MEG3)在表阿霉素抑制T细胞淋巴瘤增殖和诱导细胞凋亡中的作用及可能机制。方法实时定量PCR(RT-qPCR)检测T细胞淋巴瘤组织、临近正常组织、T细胞淋巴瘤细胞(Jurkat、CCRF-CEM、SUP-T1)、人类T细胞细胞株H9以及不同浓度表阿霉素处理的Jurkat细胞中MEG3的表达水平。将Jurkat细胞分为对照组、表阿霉素(Epirubicin)处理组、表阿霉素+pcDNA组、表阿霉素+MEG3组、表阿霉素+MEG3+miR-con组和表阿霉素+MEG3+miR-191组。采用CCK-8法、流式细胞术分别检测检测细胞活力和凋亡。蛋白质印记(Westernblot)检测B细胞淋巴瘤2(Bcl-2)和Bcl相关X蛋白(Bax)表达。双荧光素酶报告基因实验和RT-qPCR确定MEG3与miR-191靶向调控关系。结果与临近正常组织比较,T细胞淋巴瘤组织中MEG3表达明显降低(P<0.05)。与H9细胞比较,T细胞淋巴瘤细胞中MEG3表达明显降低(P<0.05)。不同浓度表阿霉素处理后Jurkat细胞中MEG3表达明显升高(P<0.05)。表阿霉素处理后Jurkat细胞存活率、Bcl-2蛋白表达明显降低,凋亡率、Bax蛋白表达明显升高(P<0.05)。过表达MEG3可提高表阿霉素对Jurkat细胞的增殖抑制和凋亡促进作用(P<0.05)。MEG3靶向miR-191并负调控其表达。过表达miR-191可逆转MEG3过表达对表阿霉素处理的Jurkat细胞增殖、凋亡以及Bcl-2和Bax蛋白表达的影响,差异有统计学意义(P<0.05)。结论LncRNA MEG3通过下调miR-191能够增强表阿霉素对T细胞淋巴瘤细胞的增殖抑制和凋亡促进作用。Objective To investigate the role of long non-coding RNA(LncRNA) matrilineal expression gene 3(MEG3) in epirubicin-induced apoptosis and anti-proliferation of human T-cell lymphoma cell, and further to explore its possible mechanism. Methods The expression levels of MEG3 in T-cell lymphoma tissues, adjacent normal tissues, T-cell lymphoma cells(Jurkat, CCRF-CEM and SUP-T1) and human T-cell cell line H9 were detected by Real-time quantitative PCR(RT-qPCR). Jurkat cells were divided into control group, Epirubicin treatment group, Epirubicin+pcDNA group, Epirubicin+MEG3 group, Epirubicin+MEG3 group, Epirubicin+MEG3+miR-con group and Epirubicin+MEG3+miR-191 group. Cell viability and apoptosis were detected by CCK-8 method and flow cytometry. The expressions of B-cell lymphoma 2(Bcl-2) and Bcl-associated X protein(Bax) were detected by Western blot.Double luciferase reporter gene experiments and RT-qPCR were performed to confirm the targeted and regulatory relationship between MEG3 and miR-191. Results Compared with adjacent normal tissues, the expression of MEG3 in T-cell lymphoma tissues was significantly reduced(P<0.05). Compared with H9 cells, the expression of MEG3 in T-cell lymphoma cells was significantly reduced(P<0.05). After treatment with different concentrations of Epirubicin, MEG3 expression in Jurkat cells was significantly increased(P<0.05). After treatment with Epirubicin, the survival rate and Bcl-2 protein expressionof Jurkat cells were significantly reduced, whereas the apoptosis rate and Bax protein expression were significantly increased(P<0.05). Overexpression of MEG3 could increase the proliferation inhibition and apoptosis promotion effect of Epirubicin on Jurkat cells(P<0.05). MEG3 targets miR-191 and negatively regulates its expression. Overexpression of miR-191 could reverse the effect of MEG3 overexpression on the proliferation and apoptosis of Epirubicin-treated Jurkat cells(P<0.05). Conclusion LncRNA MEG3 can enhance the proliferation inhibition and apoptosis promotion effect of epirubici

关 键 词：MEG3 miR-191 T细胞淋巴瘤 表阿霉素 细胞增殖 细胞凋亡 

分 类 号：R733.1[医药卫生—肿瘤]