Although it has been widely considered that all-trans retinoic acid (ATRA) is an efficient therapeutic agent for acute promyelocytic leukemia (APL), there is an urgent need for extending and examining new therapeutics in medicine. Dithiocarbamates (DTCs) are one of the recent important chemical synthetic compounds used in cancer therapy. The aim of this study was to evaluate the apoptosis-inducing effect of 2-nitro-1-phenylethylpiperidine-1-carbodithioate (2-NDC) as an active derivative from DTCs, in combination with ATRA on human APL NB4 cells. The viability of treated NB4 cells was measured by 3-(4,5-dimethyltiazol-2-yl)-2,5-diphenyltetrazolium bromide assay in various concentrations (10–120 µM). The proapoptotic effects of 2-NDC were investigated by acridine orange/ethidium bromide staining, DNA ladder formation, and flow cytometry. We also assessed the oxidative stress-inducing effect of 2-NDC and in combination with ATRA on the NB4 cells. The alteration in gene expression levels of Bax, Bcl2, and Survivin was measured through a real-time polymerase chain reaction. Furthermore, we redetected the interaction between 2-NDC and antiapoptotic proteins Bcl2 and Survivin via molecular docking. We found that 2-NDC induced apoptosis in NB4 cells in a time–dosage-dependent manner. Also, 2-NDC triggered apoptosis by expanding intracellular reactive oxygen species, combined with ATRA. Bax/Bcl2 ratio was modulated and Survivin was downregulated in NB4 cells upon 2-NDC treatment. Molecular docking studies indicated that 2-NDC binds to the baculovirus inhibitor of apoptosis protein repeat domain of Survivin and Bcl homology 3 domain of Bcl2 with various affinities. Based on the present observations, it seems that this derivative can be estimated as an appropriate candidate for future pharmaceutical evaluations.

尽管人们普遍认为全反式维甲酸（ATRA）是急性早幼粒细胞白血病（APL）的有效治疗剂，但迫切需要扩展和检查医学中的新疗法。二硫代氨基甲酸酯（DTC）是用于癌症治疗的近来重要的化学合成化合物之一。这项研究的目的是评估与DTRA结合的活性衍生物2-硝基-1-苯基乙基哌啶-1-碳二硫酸酯（2-NDC）与ATRA联合对人APL NB4细胞的凋亡诱导作用。通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四溴甲烷测定不同浓度（10-120 µM）的NB4细胞活力。通过N啶橙/溴化乙锭染色，DNA阶梯形成和流式细胞术研究了2-NDC的促凋亡作用。我们还评估了2-NDC以及与ATRA联合对NB4细胞的氧化应激诱导作用。通过实时聚合酶链反应测量Bax，Bcl2和Survivin基因表达水平的变化。此外，我们通过分子对接重新检测了2-NDC与抗凋亡蛋白Bcl2和Survivin之间的相互作用。我们发现2-NDC以时间剂量依赖性方式诱导NB4细胞凋亡。同样，2-NDC通过与ATRA结合扩展细胞内活性氧来触发细胞凋亡。2-NDC处理后，NB4细胞中的Bax / Bcl2比得到调节，而Survivin则下调。分子对接研究表明2-NDC以各种亲和力与杆状病毒抑制剂Survivin的凋亡蛋白重复结构域和Bcl2的Bcl同源性3结构域结合。