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微小RNA-21通过上调B细胞淋巴瘤-2来调节弥漫性大B细胞淋巴瘤细胞的活力和凋亡。

MicroRNA-21 regulates the viability and apoptosis of diffuse large B-cell lymphoma cells by upregulating B cell lymphoma-2.

作者信息

Liu Ke, Du Jingxia, Ruan Linhai

机构信息

Department of Hematology, The First Affiliated Hospital and College of Clinical Medicine, Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China.

Department of Pharmacology, Medical College, Henan University of Science and Technology, Luoyang, Henan 471003, P.R. China.

出版信息

Exp Ther Med. 2017 Nov;14(5):4489-4496. doi: 10.3892/etm.2017.5021. Epub 2017 Aug 24.

Abstract

Diffuse large B-cell lymphoma (DLBCL), one of the most frequently diagnosed non-Hodgkin lymphoma (NHL), is partly attributed to hereditary factors. MicroRNA-21 (miR-21) is an oncogenic substance that induces NHL and primarily targets tumor-suppressive molecules, such as B cell lymphoma-2 (Bcl-2). The present study explored whether Bcl-2, targeted by miR-21, would affect the development of NHL. Specimens were harvested from 55 patients with DLBCL who had undergone surgical treatment. Expression levels of miR-21 and Bcl-2 were evaluated through reverse transcription-quantitative polymerase chain reaction, immunohistochemistry and western blotting. Luciferase-reporter assays were performed to investigate the potential association between miR-21 and Bcl-2. MTT assays, flow cytometric analysis and caspase-3 activity assays were used to evaluate cell viability and apoptosis of DLBCL cells, respectively. Furthermore, statistical analysis was conducted using SPSS 19.0 software and the expression levels of miR-21 and Bcl-2 within DLBCL tissues were significantly upregulated when compared to those in normal tissues (P<0.01). As predicted by TargetScan, perfect base pairing was observed between the seed sequence of mature miR-21 and the 3' untranslated region of Bcl-2 mRNA. Dual luciferase reporter gene assays also revealed that miR-21 significantly facilitated the luciferase activity of Bcl-2 wild-type, with 61% upregulation (P<0.01) observed. MTT assays demonstrated that the viability of OCI-LY3 cells was decreased when cells were transfected with miR-21 inhibitor or Bcl-2 small interfering RNA and compared with those of control and negative control groups (all P<0.05). The apoptosis rate and caspase-3 activity level of the miR-21 group were 2.73±0.48 and 0.47±0.05, respectively, which were both significantly different from the groups with lower levels of miR-21 expression levels (all P<0.01). Since miR-21 may contribute to increased viability and decreased apoptosis of DLBCL cells through targeting Bcl-2, both Bcl-2 and miR-21 are likely to serve as effective targets for developing novel DLBCL treatments in the future.

摘要

弥漫性大B细胞淋巴瘤(DLBCL)是最常被诊断出的非霍奇金淋巴瘤(NHL)之一,部分归因于遗传因素。微小RNA-21(miR-21)是一种致癌物质,可诱发NHL,主要靶向肿瘤抑制分子,如B细胞淋巴瘤-2(Bcl-2)。本研究探讨了被miR-21靶向的Bcl-2是否会影响NHL的发展。从55例接受手术治疗的DLBCL患者中采集标本。通过逆转录定量聚合酶链反应、免疫组织化学和蛋白质印迹法评估miR-21和Bcl-2的表达水平。进行荧光素酶报告基因检测以研究miR-21与Bcl-2之间的潜在关联。MTT检测、流式细胞术分析和caspase-3活性检测分别用于评估DLBCL细胞的活力和凋亡。此外,使用SPSS 19.0软件进行统计分析,与正常组织相比,DLBCL组织中miR-21和Bcl-2的表达水平显著上调(P<0.01)。如TargetScan预测的那样,在成熟miR-21的种子序列与Bcl-2 mRNA的3'非翻译区之间观察到完美的碱基配对。双荧光素酶报告基因检测还显示,miR-21显著促进了Bcl-2野生型的荧光素酶活性,上调了61%(P<0.01)。MTT检测表明,当用miR-21抑制剂或Bcl-2小干扰RNA转染OCI-LY3细胞时,其活力降低,与对照组和阴性对照组相比(所有P<0.05)。miR-21组的凋亡率和caspase-3活性水平分别为2.73±0.48和0.47±0.05,与miR-21表达水平较低的组均有显著差异(所有P<0.01)。由于miR-21可能通过靶向Bcl-2导致DLBCL细胞活力增加和凋亡减少,未来Bcl-2和miR-21都可能成为开发新型DLBCL治疗方法的有效靶点。

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本文引用的文献

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