Department of Hematology, Ningbo First Hospital, Ningbo City, Zhejiang Province, 315000, China.
Department of Oncology and Hematology, Shanghai University of Medicine & Health Sciences Affiliated Zhoupu Hospital, Shanghai City, 201318, China.
Folia Histochem Cytobiol. 2020;58(3):219-226. doi: 10.5603/FHC.a2020.0018. Epub 2020 Sep 7.
microRNAs (miRNAs) are critical for tumorigenesis and progression of T-cell acute lymphoblastic leukemia (T-ALL). MiR-96-5p has been shown to play important roles in the development of many cancers, but its roles in T-ALL have yet not been studied.
miR-96-5p expression was detected in T-leukemic cells from peripheral blood of 30 patients with T-ALL using real-time quantitative PCR (RT-qPCR). TargetScan database was utilized to identify the target genes for miR-96-5p, and their target relationship was verified by western blot, dual luciferase reporter assay and RT-qPCR. The effects of miR-96-5p on the viability and proliferation of T-leukemic cells (Jurkat cells) were respectively determined using MTT and BrdU incorporation assays.
miR-96-5p presented low expression levels by qPCR in peripheral blood of T-ALL patients compared to healthy volunteers. Upregulated miR-96-5p by miR-96-5p mimic transfection markedly inhibited the viability and proliferation of Jurkat cells. Furthermore, miR-96-5p negatively regulated the expression of its target gene, HBEGF. The decreased viability and proliferation of Jurkat cells caused by miR-96-5p over-expression was suppressed after the introduction of HBEGF plasmid.
The presented study showed that upregulation of miR-96-5p inhibited the viability and proliferation of Jurkat T-leukemic cells through suppressing HBEGF expression. Our study provides a novel sight for understanding the pathological mechanism of T-ALL and suggests that miR-96-5p may be a potential biomarker for the therapy and diagnosis of T-ALL.
microRNAs (miRNAs) 在 T 细胞急性淋巴细胞白血病 (T-ALL) 的肿瘤发生和进展中起着至关重要的作用。miR-96-5p 已被证明在许多癌症的发展中发挥着重要作用，但它在 T-ALL 中的作用尚未得到研究。
采用实时定量 PCR (RT-qPCR) 检测 30 例 T-ALL 患者外周血 T 白血病细胞中 miR-96-5p 的表达。利用 TargetScan 数据库预测 miR-96-5p 的靶基因，并通过 Western blot、双荧光素酶报告基因检测和 RT-qPCR 验证其靶基因关系。分别采用 MTT 法和 BrdU 掺入法检测 miR-96-5p 对 T 白血病细胞（Jurkat 细胞）活力和增殖的影响。
与健康志愿者相比，T-ALL 患者外周血中 miR-96-5p 的 qPCR 结果显示低表达水平。miR-96-5p 模拟物转染后 miR-96-5p 的上调显著抑制 Jurkat 细胞的活力和增殖。此外，miR-96-5p 负调控其靶基因 HBEGF 的表达。HBEGF 质粒转染后，miR-96-5p 过表达引起的 Jurkat 细胞活力和增殖降低被抑制。
本研究表明，miR-96-5p 的上调通过抑制 HBEGF 表达抑制 Jurkat T 白血病细胞的活力和增殖。我们的研究为理解 T-ALL 的病理机制提供了新的视角，并表明 miR-96-5p 可能是 T-ALL 治疗和诊断的潜在生物标志物。
