Kumar Sanjeev, Tripathi Jyoti, Maurya Dharmendra K, Nuwad Jitendra, Gautam Satyendra
Food Technology Division, Bhabha Atomic Research Centre, Mumbai, 400 085 India.
Radiation Biology & Health Sciences Division, Bhabha Atomic Research Centre, Mumbai, 400 085 India.
3 Biotech. 2022 Oct;12(10):265. doi: 10.1007/s13205-022-03264-6. Epub 2022 Sep 8.
In our previous studies, a novel antimutagenic compound, 2-ethoxy-3-(3,7,11,15-tetramethylhexadec-2-ethyl) naphthaquinone-1,4-dione (ethoxy-substituted phylloquinone; ESP) from spinach was characterized and mechanism contributing to its antimutagenicity was deduced. In the current study, anti-proliferative activity of ESP was assessed in lung cancer (A549) cells using MTT [3-(4,5-dimethylthiazole-2yl)-2,5-diphenyl tetrazolium bromide], clonogenic assays and cell cycle analysis. ESP treatment showed selective cytotoxicity against lung cancer cells and no cytotoxicity in normal lung (WI38) cells. Cell cycle analysis revealed that ESP treatment arrests A549 cell population in G2-M phase. In-silico analysis indicated positive drug-likeness features of ESP. Molecular docking showed H-bonding and hydrophobic interactions between ESP and B-DNA dodecamer residues at minor groove. SWATH-MS (Sequential Window Acquisition of All Theoretical Mass Spectra) based proteomic analysis indicated down-regulation of proteins involved in EGFR signaling, NEDDylation and other metabolic pathways and up-regulation of tumor suppressor (STAT1 and NDRG1) proteins. Treatment of spinach powder with gamma radiation (5-20 kGy) from cobalt (Co-60) enhanced the extractability of ESP up to 4.4-fold at the highest dose of 20 kGy. Scanning electron microscopy of spinach powder displayed decrease in smoothness and compactness with increase in radiation dose attributing to its enhanced extractability. Increase in the extractability of ESP with increasing radiation doses as measured by fluorescence intensity and dry weight basis was strongly correlated. Nonetheless, radiation treatment did not affect the functionality of ESP in terms of anti-proliferative and antimutagenic activities. Current findings thus highlight broad spectrum bioactivity of ESP from spinach, its underlying mechanism and applicability of radiation technology in enhancing extractability.
The online version contains supplementary material available at 10.1007/s13205-022-03264-6.
在我们之前的研究中，一种来自菠菜的新型抗诱变化合物，2-乙氧基-3-(3,7,11,15-四甲基十六烷-2-乙基)萘醌-1,4-二酮（乙氧基取代的叶绿醌；ESP）被鉴定出来，并推断出其抗诱变机制。在当前研究中，使用MTT [3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑]、克隆形成试验和细胞周期分析评估了ESP在肺癌（A549）细胞中的抗增殖活性。ESP处理对肺癌细胞显示出选择性细胞毒性，而对正常肺（WI38）细胞无细胞毒性。细胞周期分析表明，ESP处理使A549细胞群体停滞在G2-M期。计算机分析表明ESP具有良好的类药性质。分子对接显示ESP与小沟处的B-DNA十二聚体残基之间存在氢键和疏水相互作用。基于SWATH-MS（所有理论质谱的顺序窗口采集）的蛋白质组学分析表明，参与表皮生长因子受体（EGFR）信号传导、NEDDylation和其他代谢途径的蛋白质下调，而肿瘤抑制蛋白（信号转导和转录激活因子1（STAT1）和N-myc下游调节基因1（NDRG1））上调。用钴（Co-60）的γ射线（5-20 kGy）处理菠菜粉，在最高剂量20 kGy时，ESP的提取率提高了4.4倍。菠菜粉的扫描电子显微镜显示，随着辐射剂量的增加，光滑度和紧实度降低，这归因于其提取率的提高。通过荧光强度和干重测量，ESP的提取率随辐射剂量的增加而增加，二者密切相关。尽管如此，辐射处理在抗增殖和抗诱变活性方面并未影响ESP的功能。因此，当前研究结果突出了菠菜中ESP的广谱生物活性、其潜在机制以及辐射技术在提高提取率方面的适用性。
在线版本包含可在10.1007/s13205-022-03264-6获取的补充材料。
