LncRNA-XIST通过抑制miR-186-5p上调PSME3表达促进急性骨髓性白血病发展的机制研究  

Promotion of acute myeloid leukemia progression by lncRNA-XIST via suppression of miR-186-5p and up-regulation of PSME3 expression

作　　者：曲海燕 姜友章 杨国鹏 孙允霄 QU Haiyan;JIANG Youzhang;YANG Guopeng;SUN Yunxiao(Department of Pediatrics,Yantai Hospital Affiliated to Binzhou Medical College,Yantai 264100,China)

机构地区：[1]滨州医学院烟台附属医院儿科,山东烟台264100

出　　处：《沈阳药科大学学报》2024年第12期1634-1643,共10页Journal of Shenyang Pharmaceutical University

摘　　要：目的探讨长链非编码RNA X染色体失活特异性转录因子(long noncoding RNA X-inactive specific transcript,lncRNA-XIST)通过抑制miR-186-5p上调PSME3表达促进急性骨髓性白血病发展的分子机制。方法采用生存曲线(Kaplan-Meier曲线)生存分析XIST表达对32例急性骨髓性白血病儿童患者生存率的影响,miRNA模拟物、miRNA抑制剂、PSME3 shRNA转染KG-1细胞后,RT-qPCR检测XIST及miR-186-5p mRNA表达水平,Western-blotting检测PSME3、cleaved-caspase 3及cleaved-caspase 8的表达水平,CCK-8及细胞凋亡实验检测KG-1细胞的活力。结果XIST在急性骨髓性白血病儿童患者临床样本及细胞系中显著表达。在KG-1细胞中,敲降XIST抑制细胞的增殖,并诱导凋亡。敲降XIST的KG-1细胞移植瘤小鼠肿瘤体积及重量显著低于对照组。双荧光素酶报告基因检测结果显示miR-186-5p与XIST相互作用,机制研究发现XIST通过内源性竞争海绵吸附miR-186-5p影响其下游靶基因PSME3的表达。结论XIST在急性骨髓性白血病中可通过海绵吸附miR-186-5p调控PSME3表达,XIST的异常表达可作为急性骨髓性白血病诊断及治疗的作用靶点。Objective To investigate the mechanism of long intergenic noncoding RNA X-inactive specific transcript(lncRNA-XIST)promoted acute myeloid leukemia(AML)progression by suppressing miR-186-5p.Methods Kaplan-meier was used to analyze the correlation between XIST expression and survival in 32 AML patients.KG-1 cells were tranfected with miRNA mimic,miRNA inhibitor or PSME3 shRNAs.Subsequently,the mRNA expression levels of XIST and miR-186-5p were detected by quantitative real-time PCR.The protein expression levels of PSME3,cleaved-caspase 3 and cleaved-caspase 8 were detected by Western-blotting.Cell viability detected by CCK-8 assay and flow cytometry.Results XIST were significantly expressedby AML patients and AML cell lines.Knocking down XIST inhibited the proliferation and promoted the apoptosis of KG-1 cells.Moreover,when KG-1 cells with XIST knockdown were transplanted into nude mice,tumor formation and progression were remarkably suppressed.Dual-luciferase reporter assay showed that miR-186-5p targeted XIST.The mechanism investigation showd that XIST endogenously competed sponge miR-186-5p regulate PSME3 expression.Conclusion XIST endogenously competed as a sponge for miR-186-5p to regulate PSME3 expression in AML,and the aberrant expression of XIST is a potential molecular target for AML diagnosis and therapy.

关 键 词：急性骨髓性白血病 XIST miR-186-5p PSME3 

分 类 号：R733.71[医药卫生—肿瘤]