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Open AccessArticle

Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves Salix safsaf

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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
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Department of Basic Medical Sciences, Kulliyyah (College) of Medicine, International Islamic University Malaysia, Kuantan 25200, Pahang, Malaysia
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Toxicology Centre, University of Saskatchewan, Saskatoon, SK S7N 5B3, Canada
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Department of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia
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Department of Pharmacognosy, Faculty of Pharmacy, Mansoura University, Mansoura 35516, Egypt
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Vitiligo Research Chair, College of Medicine, King Saud University, Riyadh 11451, Saudi Arabia
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Biochemistry Department, Faculty of Agriculture, Cairo University, Giza 12613, Egypt
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Zoology Department, College of Sciences, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia
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Department of Dermatology, College of Medicine, P.O. Box 240997, King Saud University, Riyadh 11322, Saudi Arabia
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School of Environment and Sustainability, University of Saskatchewan, Saskatoon, SK S7N 5B3, Canada
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Department of Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, SK S7N 5B4, Canada
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Department of Environmental Science, Baylor University, Waco, TX 76798-7266 (254), USA
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Authors to whom correspondence should be addressed.
Plants 2020, 9(10), 1295; https://doi.org/10.3390/plants9101295
Received: 3 September 2020 / Revised: 23 September 2020 / Accepted: 28 September 2020 / Published: 30 September 2020
Organic fractions and extracts of willow (Salix safsaf) 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 (CHCl3) 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 H2O2 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. View Full-Text
Keywords: Salix safsaf; cytotoxicity; polyphenols; flavonoids; mass spectrometry; apoptosis; natural products Salix safsaf; cytotoxicity; polyphenols; flavonoids; mass spectrometry; apoptosis; natural products
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MDPI and ACS Style

Aboul-Soud, M.A.M.; Ashour, A.E.; Challis, J.K.; Ahmed, A.F.; Kumar, A.; Nassrallah, A.; Alahmari, T.A.; Saquib, Q.; Siddiqui, M.A.; Al-Sheikh, Y.; El-Shemy, H.A.; Aboul-Enein, A.M.; Alghamdi, K.M.; Jones, P.D.; Giesy, J.P. Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves Salix safsaf. Plants 2020, 9, 1295. https://doi.org/10.3390/plants9101295

AMA Style

Aboul-Soud MAM, Ashour AE, Challis JK, Ahmed AF, Kumar A, Nassrallah A, Alahmari TA, Saquib Q, Siddiqui MA, Al-Sheikh Y, El-Shemy HA, Aboul-Enein AM, Alghamdi KM, Jones PD, Giesy JP. Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves Salix safsaf. Plants. 2020; 9(10):1295. https://doi.org/10.3390/plants9101295

Chicago/Turabian Style

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. 2020. "Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves Salix safsaf" Plants 9, no. 10: 1295. https://doi.org/10.3390/plants9101295

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