Niakani Maryam, Majd Ahmad, Pakzad Parviz, Malekinejad Hassan
Department of Cellular and Molecular Biology, Faculty of Biological Sciences, Tehran North Branch, Islamic Azad University, Tehran, I.R. Iran.
Department of Microbiology, Faculty of Biological Sciences, Tehran North Branch, Islamic Azad University, Tehran, I.R. Iran.
Res Pharm Sci. 2020 Dec 30;16(1):26-34. doi: 10.4103/1735-5362.305186. eCollection 2021 Feb.
Chronic myeloid leukemia (CML) as a myeloproliferative disease is characterized by increased cellularity of bone marrow. Implementing the latest treatment protocols is currently accompanied by serious and life-threatening side effects. There are worldwide attempts to find new effective and potent therapeutic agents with minimal side effects on CML patients. This study was carried out to discover the potential antiproliferative and apoptotic effects of naturally produced prodigiosin (PDG) on K562 cells as an accepted model of CML.
The anti-proliferative effect of PDG was measured by MTT assay. To highlight the mechanism of cytotoxicity, the apoptotic cell death pathway was investigated by morphological and biochemical assessments. The dual acridine orange/ethidium bromide staining technique and western blotting method were applied to assess the mechanism of the potential apoptotic impact of PDG on K562 cells.
FINDINGS/RESULTS: PDG-induced time- and concentration-dependent anti-proliferative effects were revealed with an estimated IC value of 54.06 μM. The highest cell viability reduction (60%) was recorded in cells, which were exposed to 100 μM concentration. Further assays demonstrated that in the dual acridine orange/ethidium bromide staining method the cell population in the late apoptosis phase was increased in a concentration-dependent manner, which was confirmed with remarkable DNA fragmentation.
We found that the PDG-induced apoptosis in K562 cells is mediated through the caspase-3 activation both in mRNA and protein levels. Our results suggest that PDG could be a potent compound for further pharmacokinetic and pharmacodynamics studies in the model of CML.
慢性粒细胞白血病（CML）作为一种骨髓增殖性疾病，其特征为骨髓细胞增多。目前实施最新治疗方案会伴随严重且危及生命的副作用。全球都在尝试寻找对CML患者副作用最小的新型有效治疗药物。本研究旨在探索天然产生的灵菌红素（PDG）对作为CML公认模型的K562细胞的潜在抗增殖和凋亡作用。
通过MTT法检测PDG的抗增殖作用。为突出细胞毒性机制，通过形态学和生化评估研究凋亡细胞死亡途径。应用双吖啶橙/溴化乙锭染色技术和蛋白质印迹法评估PDG对K562细胞潜在凋亡影响的机制。
PDG诱导的抗增殖作用呈时间和浓度依赖性，估计IC值为54.06μM。在暴露于100μM浓度的细胞中，细胞活力降低最多（60%）。进一步检测表明，在双吖啶橙/溴化乙锭染色法中，晚期凋亡阶段的细胞群体呈浓度依赖性增加，这通过显著的DNA片段化得到证实。
我们发现PDG诱导K562细胞凋亡是通过在mRNA和蛋白质水平上激活半胱天冬酶-3介导的。我们的结果表明，PDG可能是一种在CML模型中进行进一步药代动力学和药效学研究的有效化合物。
