Medical Biology Department, Ege University Medicine Faculty, Izmir, Turkey.
Bosn J Basic Med Sci. 2018 May 20;18(2):141-149. doi: 10.17305/bjbms.2017.2457.
Matrine, a natural product extracted from the root of Sophora flavescens, is a promising alternative drug in different types of cancer. Here, we aimed to investigate the therapeutic effects and underlying molecular mechanisms of matrine on human acute lymphoblastic leukemia (ALL) cell line, CCRF-CEM. Cell viability and IC50 values were determined by WST-1 cell cytotoxicity assay. Cell cycle distribution and apoptosis rates were analyzed by flow cytometry. Expression patterns of 44 selected miRNAs and 44 RNAs were analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) using the Applied Biosystems 7500 Fast Real-Time PCR System. Matrine inhibited cell viability and induced apoptosis of CCRF-CEM cells in a dose-dependent manner. Cell cycle analysis demonstrated that matrine-treated CCRF-CEM cells significantly accumulated in the G0/G1 phase compared with the untreated control cells. hsa-miR-376b-3p (-37.09 fold, p = 0.008) and hsa-miR-106b-3p (-16.67 fold, p = 0.028) expressions were decreased, whereas IL6 (95.47 fold, p = 0.000011) and CDKN1A (140.03 fold, p = 0.000159) expressions were increased after matrine treatment. Our results suggest that the downregulation of hsa-miR-106b-3p leads to the upregulation of target p21 gene, CDKN1A, and plays a critical role in the cell cycle progression by arresting matrine-treated cells in the G0/G1 phase.
苦参碱是从苦参根中提取的天然产物,是一种有前途的治疗多种癌症的药物。在这里,我们旨在研究苦参碱对人急性淋巴细胞白血病(ALL)细胞系 CCRF-CEM 的治疗效果和潜在的分子机制。通过 WST-1 细胞细胞毒性测定法测定细胞活力和 IC50 值。通过流式细胞术分析细胞周期分布和凋亡率。使用 Applied Biosystems 7500 Fast Real-Time PCR System 通过定量逆转录聚合酶链反应(qRT-PCR)分析 44 种选定的 miRNA 和 44 种 RNA 的表达模式。苦参碱呈剂量依赖性抑制 CCRF-CEM 细胞的活力并诱导其凋亡。细胞周期分析表明,苦参碱处理的 CCRF-CEM 细胞与未处理的对照细胞相比,显著在 G0/G1 期积累。hsa-miR-376b-3p(-37.09 倍,p=0.008)和 hsa-miR-106b-3p(-16.67 倍,p=0.028)的表达降低,而 IL6(95.47 倍,p=0.000011)和 CDKN1A(140.03 倍,p=0.000159)的表达增加。我们的研究结果表明,hsa-miR-106b-3p 的下调导致靶基因 p21 的上调,CDKN1A,通过阻止苦参碱处理的细胞进入 G0/G1 期,在细胞周期进展中发挥关键作用。