Zhou Ling, Bu Yunwen, Liang Yanyan, Zhang Fang, Zhang Haiguo, Li Shumei
Department of Hematology, The First People's Hospital, Jining, Shandong, China (mainland).
Med Sci Monit. 2016 Oct 31;22:4114-4120. doi: 10.12659/msm.897306.
BACKGROUND This study was designed to explore the molecular mechanism underlying the effect of cellular miRNAs and EBV miRNA upon the expression of targets such as PTEN, and their involvement in the pathogenesis of Burkitt lymphoma. MATERIAL AND METHODS In this study, we examined several differentially expressed cellular miRNAs in EBV-positive versus EBV-negative Burkett lymphoma tissue samples, and confirmed PTEN as targets of cellular miR-142 by using a bioinformatics tool, luciferase reporter system, oligo transfection, real-time PCR, and Western blot analysis. RESULTS We further confirmed the binding site of miR-142 in the 3'UTR of the target genes, and established the negative regulatory relationship between miRNA and mRNAs with luciferase activity assay. To verify the regulatory relationship between the miRNAs and PTEN, we evaluated the expression of PTEN in the tissue samples, and found that PTEN was downregulated in EBV- positive Burkett lymphoma. Additionally, lymphoma cells were transfected with EBV-BART-6-3p and miR-142 and we found that EBV-BART-6-3p and miR-142 synergistically reduced expression of IL-6R and PTEN. Furthermore, we also examined viability of the cells in each treatment group, and showed that EBV-BART-6-3p and miR-142 synergistically promoted proliferation of the cells. CONCLUSIONS These findings improve our knowledge about the role of miR-142/EBV-BART-6-3p and their target, PTEN, in the development of Burkett lymphoma; they could be novel therapeutic targets for the treatment of EBV-positive Burkett lymphoma.
背景 本研究旨在探索细胞微小RNA(miRNA)和EB病毒(EBV)miRNA对诸如PTEN等靶标表达产生影响的分子机制,以及它们在伯基特淋巴瘤发病机制中的作用。
材料与方法 在本研究中,我们检测了EBV阳性与EBV阴性伯基特淋巴瘤组织样本中几种差异表达的细胞miRNA,并通过生物信息学工具、荧光素酶报告系统、寡核苷酸转染、实时聚合酶链反应(PCR)和蛋白质免疫印迹分析,证实PTEN是细胞miR-142的靶标。
结果 我们进一步证实了miR-142在靶基因3'非翻译区(UTR)的结合位点,并通过荧光素酶活性测定建立了miRNA与mRNA之间的负调控关系。为了验证miRNA与PTEN之间的调控关系,我们评估了组织样本中PTEN的表达,发现PTEN在EBV阳性伯基特淋巴瘤中表达下调。此外,用EBV-BART-6-3p和miR-142转染淋巴瘤细胞,我们发现EBV-BART-6-3p和miR-142协同降低白细胞介素6受体(IL-6R)和PTEN的表达。此外,我们还检测了各治疗组细胞的活力,结果显示EBV-BART-6-3p和miR-142协同促进细胞增殖。
结论 这些发现增进了我们对miR-142/EBV-BART-6-3p及其靶标PTEN在伯基特淋巴瘤发生发展中作用的认识;它们可能是治疗EBV阳性伯基特淋巴瘤的新型治疗靶点。