Aboul-Soud Mourad A M, Ashour Abdelkader E, Challis Jonathan K, Ahmed Atallah F, Kumar Ashok, Nassrallah Amr, Alahmari Tariq A, Saquib Quaiser, Siddiqui Maqsood A, Al-Sheikh Yazeed, El-Shemy Hany A, Aboul-Enein Ahmed M, Alghamdi Khalid M, Jones Paul D, Giesy John P
Chair of Medical and Molecular Genetics Research, Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Saud University, P.O. Box 10219, Riyadh 11433, Saudi Arabia.
Department of Basic Medical Sciences, Kulliyyah (College) of Medicine, International Islamic University Malaysia, Kuantan 25200, Pahang, Malaysia.
Plants (Basel). 2020 Sep 30;9(10):1295. doi: 10.3390/plants9101295.
Organic fractions and extracts of willow () leaves, produced by sequential solvent extraction as well as infusion and decoction, exhibited anticancer potencies in four cancerous cell lines, including breast (MCF-7), colorectal (HCT-116), cervical (HeLa) and liver (HepG2). Results of the MTT assay revealed that chloroform (CHCl) and ethyl acetate (EtOAc)-soluble fractions exhibited specific anticancer activities as marginal toxicities were observed against two non-cancerous control cell lines (BJ-1 and MCF-12). Ultra-high-resolution mass spectrometry Q-Exactive™ HF Hybrid Quadrupole-Orbitrap™ coupled with liquid chromatography (UHPLC) indicated that both extracts are enriched in features belonging to major phenolic and purine derivatives. Fluorescence-activated cell sorter analysis (FACS), employing annexin V-FITC/PI double staining indicated that the observed cytotoxic potency was mediated via apoptosis. FACS analysis, monitoring the increase in fluorescence signal, associated with oxidation of DCFH to DCF, indicated that the mechanism of apoptosis is independent of reactive oxygen species (ROS). Results of immunoblotting and RT-qPCR assays showed that treatment with organic fractions under investigation resulted in significant up-regulation of pro-apoptotic protein and mRNA markers for Caspase-3, p53 and Bax, whereas it resulted in a significant reduction in amounts of both protein and mRNA of the anti-apoptotic marker Bcl-2. FACS analysis also indicated that pre-treatment and co-treatment of human amniotic epithelial (WISH) cells exposed to the ROS HO with EtOAc fraction provide a cytoprotective and antioxidant capacity against generated oxidative stress. In conclusion, our findings highlight the importance of natural phenolic and flavonoid compounds with unparalleled and unique antioxidant and anticancer properties.
通过连续溶剂萃取以及浸泡和煎煮法制备的柳树（ ）叶的有机馏分和提取物，在四种癌细胞系中表现出抗癌效力，这四种癌细胞系包括乳腺癌（MCF-7）、结肠直肠癌（HCT-116）、宫颈癌（HeLa）和肝癌（HepG2）。MTT 分析结果显示，氯仿（CHCl）和乙酸乙酯（EtOAc）可溶馏分表现出特定的抗癌活性，因为在两种非癌对照细胞系（BJ-1 和 MCF-12）中观察到了轻微的毒性。超高分辨率质谱 Q-Exactive™ HF 混合型四极杆-轨道阱™ 与液相色谱（UHPLC）联用表明，两种提取物都富含属于主要酚类和嘌呤衍生物的特征物。采用膜联蛋白 V-FITC/PI 双重染色的荧光激活细胞分选分析（FACS）表明，观察到的细胞毒性效力是通过凋亡介导的。监测与 DCFH 氧化为 DCF 相关的荧光信号增加的 FACS 分析表明，凋亡机制独立于活性氧（ROS）。免疫印迹和 RT-qPCR 分析结果表明，用所研究的有机馏分处理导致促凋亡蛋白以及半胱天冬酶-3、p53 和 Bax 的 mRNA 标志物显著上调，而抗凋亡标志物 Bcl-2 的蛋白和 mRNA 量则显著减少。FACS 分析还表明，用 EtOAc 馏分对暴露于 ROS HO 的人羊膜上皮（WISH）细胞进行预处理和共处理，可提供针对所产生氧化应激的细胞保护和抗氧化能力。总之，我们的研究结果突出了具有无与伦比和独特抗氧化及抗癌特性的天然酚类和黄酮类化合物的重要性。
