Department of Hematopathology, Yantaishan Hospital, Yantai, China.
Eur Rev Med Pharmacol Sci. 2020 Dec;24(23):12063-12072. doi: 10.26355/eurrev_202012_23995.
The aim of this study was to explore the influences of micro ribonucleic acid (miR)-150 on the proliferation and apoptosis of mantle-cell lymphoma (MCL) cells and to investigate the potential underlying mechanism.
Differentially expressed miRNAs in MCL tissues were excavated via microarray analysis of miRNA expression profiles. Subsequently, the expression of miRNAs were verified by quantitative Reverse Transcription-Polymerase Chain Reaction (RT-qPCR). The influence of miRNA expression on the survival of patients was detected based on clinical data. Besides, the potential targets of miRNAs were determined using Luciferase reporter gene assay combined with qRT-PCR and Western blotting. Primary tumor cells were extracted, and the influences of miR-150 expression on cell proliferation were detected via Cell Counting Kit (CCK)-8 assay and 5-ethynyl-2'-deoxyuridine (EdU) staining assay. Finally, Western blotting and flow cytometry were performed to explore the impact of miR-150 on the apoptosis of primary tumor cells.
Microarray analysis of miRNA expression profiles and RT-qPCR verified that the expression levels of hsa-miR-486, hsa-miR-4746, and hsa-miR-3158 rose considerably in MCL tissues, while those of hsa-miR-29b-3p, hsa-miR-150, and hsa-miR-142-5p remarkably declined. According to the results of survival analysis, the survival time was notably prolonged in patients with higher expression levels of miR-150 and miR-486, especially in those with higher expression level of miR-150. Luciferase reporter gene assay and RT-qPCR and Western blotting results demonstrated that miR-150 negatively regulated the expression level of MET. Subsequent CCK-8 assay and EdU staining results revealed that up-regulation of miR-150 significantly suppressed the proliferation of primary MCL cells. Finally, Western blotting and flow cytometry found that increased expression of MET remarkably facilitated the apoptosis of primary MCL cells.
MiR-150 inhibits the proliferation and promotes the apoptosis of MCL cells by negatively regulating MET expression.
本研究旨在探讨微小 RNA（miR）-150 对套细胞淋巴瘤（MCL）细胞增殖和凋亡的影响，并探讨潜在的作用机制。
通过 miRNA 表达谱的微阵列分析挖掘出 MCL 组织中差异表达的 miRNAs，然后通过定量逆转录-聚合酶链反应（RT-qPCR）验证 miRNA 的表达。根据临床资料检测 miRNA 表达对患者生存的影响。此外，通过荧光素酶报告基因检测联合 qRT-PCR 和 Western blot 确定 miRNA 的潜在靶标。提取原代肿瘤细胞，通过 CCK-8 检测和 5-乙炔基-2'-脱氧尿苷（EdU）染色检测 miR-150 表达对细胞增殖的影响。最后，通过 Western blot 和流式细胞术探讨 miR-150 对原代肿瘤细胞凋亡的影响。
miRNA 表达谱的微阵列分析和 RT-qPCR 验证 hsa-miR-486、hsa-miR-4746 和 hsa-miR-3158 在 MCL 组织中的表达水平显著升高，而 hsa-miR-29b-3p、hsa-miR-150 和 hsa-miR-142-5p 的表达水平显著降低。根据生存分析结果，miR-150 和 miR-486 表达水平较高的患者的生存时间明显延长，尤其是 miR-150 表达水平较高的患者。荧光素酶报告基因检测以及 RT-qPCR 和 Western blot 结果表明，miR-150 负调控 MET 的表达水平。随后的 CCK-8 检测和 EdU 染色结果表明，miR-150 的上调显著抑制原代 MCL 细胞的增殖。最后，Western blot 和流式细胞术发现 MET 表达增加显著促进原代 MCL 细胞的凋亡。
miR-150 通过负调控 MET 的表达抑制 MCL 细胞的增殖并促进其凋亡。
