Wang Hui, Chinnathambi Arunachalam, Alahmadi Tahani Awad, Alharbi Sulaiman Ali, Veeraraghavan Vishnu Priya, Krishna Mohan Surapaneni, Hussain Sardar, Ramamoorthy Kavitha, Rengarajan Thamaraiselvan
Department of Hematology, Shaanxi Provincial People's Hospital, Xi'an, China.
Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, Saudi Arabia.
J Biochem Mol Toxicol. 2021 Jun;35(6):1-10. doi: 10.1002/jbt.22758. Epub 2021 Mar 16.
Among cancers, leukemia is a multistep progression that involves genetic modifications of normal hematopoietic progenitor cells to cancerous cells. In recent times, leukemia cases and their mortality rate have increased rapidly. Therefore, the immense need for a therapeutic approach is crucial that can control this type of cancer. Phyllanthin is a lignan compound constituent from the Phyllanthus species and has numerous beneficial effects as a dietary component. The present study aims to determine the impact of phyllanthin on the MOLT-4 cytotoxic effect. MOLT-4 cells and MS-5 cells were cultured at different concentrations of phyllanthin (5, 10, 25, 50, 75, and 100 μM/ml), and the viability was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. The level of reactive oxygen species, the membrane potential of mitochondria, apoptosis by 2',7'-dichlorofluorescin-diacetate (DCF-DA), rhodamine, acridine orange (AO)/ethidium bromide (EB), 4',6-diamidino-2-phenylindole (DAPI)/propidium iodide (PI) staining, gene expression of signaling molecules, and protein levels were assessed by reverse-transcription polymerase chain reaction and western blot analysis. Phyllanthin did not show toxicity toward MS-5 cells and significantly decreased the cell viability of MOLT-4 cells with an IC value of 25 µM/ml. Also, phyllanthin induced the production of reactive oxygen species and led to the loss of mitochondrial membrane potential. AO/EB and DAPI/PI staining fluorescent image confirmed the induction of apoptosis by phyllanthin treatment. The messenger RNA (mRNA) expression of cell cycle regulator cyclin D1, antiapoptotic gene Bcl-2, NF-κB, and TNF-α decreased, but the proapoptotic Bax mRNA expression was increased. The phosphorylated protein levels of p-PI3K1/2, p-ERK1/2, and p-AKT were decreased, whereas the levels of p-p38 and p-JNKT1/2 increased. Our results confirmed that phyllanthin inhibits the MOLT-4 cells, increases apoptosis, and inhibits MOLT-4 migration and cell invasion. Therefore, phyllanthin can be used as a potential target for leukemia treatment.
在癌症中，白血病是一个多步骤的进展过程，涉及正常造血祖细胞向癌细胞的基因修饰。近年来，白血病病例及其死亡率迅速上升。因此，迫切需要一种能够控制这类癌症的治疗方法。叶下珠素是一种来自叶下珠属植物的木脂素类化合物成分，作为一种饮食成分具有多种有益作用。本研究旨在确定叶下珠素对MOLT-4细胞的细胞毒性作用。将MOLT-4细胞和MS-5细胞在不同浓度的叶下珠素（5、10、25、50、75和100μM/ml）中培养，并通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法评估细胞活力。通过逆转录聚合酶链反应和蛋白质印迹分析评估活性氧水平、线粒体膜电位、2',7'-二氯荧光素二乙酸酯（DCF-DA）诱导的凋亡、罗丹明、吖啶橙（AO）/溴化乙锭（EB）、4',6-二脒基-2-苯基吲哚（DAPI）/碘化丙啶（PI）染色、信号分子的基因表达以及蛋白质水平。叶下珠素对MS-5细胞未显示出毒性，并且以25μM/ml的IC值显著降低了MOLT-4细胞的活力。此外，叶下珠素诱导活性氧的产生并导致线粒体膜电位的丧失。AO/EB和DAPI/PI染色荧光图像证实了叶下珠素处理诱导了细胞凋亡。细胞周期调节因子细胞周期蛋白D1、抗凋亡基因Bcl-2、NF-κB和TNF-α的信使核糖核酸（mRNA）表达降低，但促凋亡的Bax mRNA表达增加。p-PI3K1/2、p-ERK1/2和p-AKT的磷酸化蛋白水平降低，而p-p38和p-JNKT-1/2的水平升高。我们的结果证实叶下珠素抑制MOLT-4细胞生长，增加细胞凋亡，并抑制MOLT-4细胞迁移和侵袭。因此，叶下珠素可作为白血病治疗的潜在靶点。
