Mohammed Mohammed S, Al-Taee Maha F, Al-Shammari Ahmed Majeed
Kufa Institute, Al-Furat Al-Awsat University, Iraq.
Biotechnology Department, college of Science, University of Baghdad, Baghdad, Iraq.
Int J Mol Cell Med. 2019 Summer;8(3):211-223. doi: 10.22088/IJMCM.BUMS.8.3.211.
Hematological malignancies remain one of the leading causes of death worldwide despite advances in cancer therapeutics. Newcastle disease virus (NDV) is a member of that elicits considerable interest as an anticancer agent because it can replicate up to 10 000 times faster in human cancer cells than in most normal cancer cells. Several NDV strains reportedly induce the cytolysis of cancerous cell lines. The attenuated Iraqi strain (AMHA1) of NDV is a novel oncolytic agent with promising antitumor characteristics, including apoptosis induction. This study aimed to evaluate the ability of the AMHA1 NDV strain to induce apoptotic cell death in hematological tumors through caspase-dependent or independent apoptotic pathways. The cytolytic effects of AMHA1 NDV strains of different multiplicity of infection (MOIs) (20, 15,10, 5, 3, 1, 0.5, and 0.1 )and exposure for all hematological malignancy cell lines (human non-Hodgkin lymphoma SR and human multiple myeloma (COLO 677) and human monocytic leukemia THP1) have been determined through a microtetrazolium (MTT) assay. Propidium iodide and acridine orange (AO/PI) double staining were used to examine the ability of attenuated NDV strain to induce apoptosis in infected cells under a fluorescence microscope and to quantify the percentage of apoptosis induction. Quantitative immunocytochemistry assay was further used to study the caspase-dependent and independent protein expression levels in infected and control cells. Cells treated with NDV strains showed a higher cell-death percentage than untreated cells as quantified by the MTT assay. AO/PI results revealed that NDV exerted a powerful and significant effect on apoptosis induction (P<0.0001) in the human cancer cell lines tested in comparison with control cells. Immunocytochemistry in AMHA1 NDV- infected human hematological cell lines revealed a remarkable increase in the expression of caspase 8, 9 (dependent pathway), apoptosis-inducing factor, and endonuclease G (independent pathway) in comparison with untreated cells. This study demonstrated the role of the Iraqi NDV strain in inducing apoptosis through dependent and independent pathways in cancer cells and thus its high potential as an antitumor agent.
尽管癌症治疗取得了进展，但血液系统恶性肿瘤仍然是全球主要的死亡原因之一。新城疫病毒（NDV）是其中一员，作为一种抗癌剂引起了人们的极大兴趣，因为它在人类癌细胞中的复制速度比大多数正常细胞快10000倍。据报道，几种NDV毒株可诱导癌细胞系的细胞溶解。NDV的减毒伊拉克毒株（AMHA1）是一种新型溶瘤剂，具有包括诱导凋亡在内的有前景的抗肿瘤特性。本研究旨在评估AMHA1 NDV毒株通过半胱天冬酶依赖性或非依赖性凋亡途径诱导血液肿瘤细胞凋亡的能力。通过微量四氮唑蓝（MTT）试验测定了不同感染复数（MOI）（20、15、10、5、3、1、0.5和0.1）的AMHA1 NDV毒株对所有血液系统恶性肿瘤细胞系（人非霍奇金淋巴瘤SR、人多发性骨髓瘤（COLO 677）和人单核细胞白血病THP1）的细胞溶解作用以及暴露后的影响。碘化丙啶和吖啶橙（AO/PI）双重染色用于在荧光显微镜下检查减毒NDV毒株诱导感染细胞凋亡的能力，并量化凋亡诱导百分比。进一步采用定量免疫细胞化学分析来研究感染细胞和对照细胞中半胱天冬酶依赖性和非依赖性蛋白表达水平。通过MTT试验定量分析，用NDV毒株处理的细胞显示出比未处理细胞更高的细胞死亡百分比。AO/PI结果显示，与对照细胞相比，NDV对所测试的人类癌细胞系的凋亡诱导具有强大且显著的作用（P<0.0001）。对AMHA1 NDV感染的人类血液细胞系进行免疫细胞化学分析显示，与未处理细胞相比，半胱天冬酶8、9（依赖性途径）、凋亡诱导因子和核酸内切酶G（非依赖性途径）的表达显著增加。本研究证明了伊拉克NDV毒株在癌细胞中通过依赖性和非依赖性途径诱导凋亡的作用，因此其作为抗肿瘤剂具有很高的潜力。
