Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves <i>Salix safsaf</i>
Organic fractions and extracts of willow (<i>Salix safsaf</i>) 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 (HepG...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-09-01
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| Series: | Plants |
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| Online Access: | https://www.mdpi.com/2223-7747/9/10/1295 |
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| author | Mourad A. M. Aboul-Soud Abdelkader E. Ashour Jonathan K. Challis Atallah F. Ahmed Ashok Kumar Amr Nassrallah Tariq A. Alahmari Quaiser Saquib Maqsood A. Siddiqui Yazeed Al-Sheikh Hany A. El-Shemy Ahmed M. Aboul-Enein Khalid M. Alghamdi Paul D. Jones John P. Giesy |
| author_facet | Mourad A. M. Aboul-Soud Abdelkader E. Ashour Jonathan K. Challis Atallah F. Ahmed Ashok Kumar Amr Nassrallah Tariq A. Alahmari Quaiser Saquib Maqsood A. Siddiqui Yazeed Al-Sheikh Hany A. El-Shemy Ahmed M. Aboul-Enein Khalid M. Alghamdi Paul D. Jones John P. Giesy |
| author_sort | Mourad A. M. Aboul-Soud |
| collection | DOAJ |
| description | Organic fractions and extracts of willow (<i>Salix safsaf</i>) 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<sub>3</sub>) 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 H<sub>2</sub>O<sub>2</sub> 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. |
| first_indexed | 2024-03-10T15:55:42Z |
| format | Article |
| id | doaj.art-38cc45ec1ffb48c281c4965896b2939a |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T15:55:42Z |
| publishDate | 2020-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj.art-38cc45ec1ffb48c281c4965896b2939a2023-11-20T15:41:54ZengMDPI AGPlants2223-77472020-09-01910129510.3390/plants9101295Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves <i>Salix safsaf</i>Mourad A. M. Aboul-Soud0Abdelkader E. Ashour1Jonathan K. Challis2Atallah F. Ahmed3Ashok Kumar4Amr Nassrallah5Tariq A. Alahmari6Quaiser Saquib7Maqsood A. Siddiqui8Yazeed Al-Sheikh9Hany A. El-Shemy10Ahmed M. Aboul-Enein11Khalid M. Alghamdi12Paul D. Jones13John P. Giesy14Chair 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 ArabiaDepartment of Basic Medical Sciences, Kulliyyah (College) of Medicine, International Islamic University Malaysia, Kuantan 25200, Pahang, MalaysiaToxicology Centre, University of Saskatchewan, Saskatoon, SK S7N 5B3, CanadaDepartment of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh 11451, Saudi ArabiaVitiligo Research Chair, College of Medicine, King Saud University, Riyadh 11451, Saudi ArabiaBiochemistry Department, Faculty of Agriculture, Cairo University, Giza 12613, EgyptChair 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 ArabiaZoology Department, College of Sciences, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi ArabiaZoology Department, College of Sciences, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi ArabiaChair 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 ArabiaBiochemistry Department, Faculty of Agriculture, Cairo University, Giza 12613, EgyptBiochemistry Department, Faculty of Agriculture, Cairo University, Giza 12613, EgyptVitiligo Research Chair, College of Medicine, King Saud University, Riyadh 11451, Saudi ArabiaToxicology Centre, University of Saskatchewan, Saskatoon, SK S7N 5B3, CanadaToxicology Centre, University of Saskatchewan, Saskatoon, SK S7N 5B3, CanadaOrganic fractions and extracts of willow (<i>Salix safsaf</i>) 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<sub>3</sub>) 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 H<sub>2</sub>O<sub>2</sub> 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.https://www.mdpi.com/2223-7747/9/10/1295<i>Salix safsaf</i>cytotoxicitypolyphenolsflavonoidsmass spectrometryapoptosis |
| spellingShingle | Mourad A. M. Aboul-Soud Abdelkader E. Ashour Jonathan K. Challis Atallah F. Ahmed Ashok Kumar Amr Nassrallah Tariq A. Alahmari Quaiser Saquib Maqsood A. Siddiqui Yazeed Al-Sheikh Hany A. El-Shemy Ahmed M. Aboul-Enein Khalid M. Alghamdi Paul D. Jones John P. Giesy Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves <i>Salix safsaf</i> Plants <i>Salix safsaf</i> cytotoxicity polyphenols flavonoids mass spectrometry apoptosis |
| title | Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves <i>Salix safsaf</i> |
| title_full | Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves <i>Salix safsaf</i> |
| title_fullStr | Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves <i>Salix safsaf</i> |
| title_full_unstemmed | Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves <i>Salix safsaf</i> |
| title_short | Biochemical and Molecular Investigation of In Vitro Antioxidant and Anticancer Activity Spectrum of Crude Extracts of Willow Leaves <i>Salix safsaf</i> |
| title_sort | biochemical and molecular investigation of in vitro antioxidant and anticancer activity spectrum of crude extracts of willow leaves i salix safsaf i |
| topic | <i>Salix safsaf</i> cytotoxicity polyphenols flavonoids mass spectrometry apoptosis |
| url | https://www.mdpi.com/2223-7747/9/10/1295 |
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