Phytochemical screening and in vitro anticancer activity of ethyl acetate fraction of seagrass Halodule uninervis from Mandapam coastal region, Rameswaram, Gulf of Mannar, India
Abstract
Seagrasses are one of the groups of marine angiosperms widely distributed all over the coastal regions. The seagrass ecosystem is very productive and traditionally used for a variety of therapeutic purposes. The present study aimed to investigate the in-vitro anticancer activity against malignant melanoma (A375), lung carcinoma (A549), cervix adenocarcinoma (HeLa) and colorectal adenocarcinoma (HT29) cell lines and phytochemical screening of ethyl acetate fraction of Halodule uninervis (EAF) from Mandapam coastal region, Rameswaram, Gulf of Mannar, India. The qualitative phytochemical analysis of EAF was performed as followed by standard methods. EAF showed the presence of alkaloids, steroids, terpenoids, flavonoids, phenols and quinones. The total flavonoid and phenolic contents of EAF were determined using the spectrophotometric method. The total flavonoid and phenolic contents showed 67.33 ± 1.52 mg QE/g dried fraction and 87.75 ± 1.39 mg GAE/g dried fraction, respectively. Gas Chromatography-Mass Spectrometry (GC-MS) analysis of EAF shows the presence of 23 phytochemical compounds. The in-vitro anticancer activity of EAF was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, AO/EtBr staining and DNA fragmentation assay for 24 hours treatment. The IC50 values of EAF showed 91.64 ± 1.61 μg, 269.16 ± 5.17 μg, 845.16 ± 36.09 μg and >1000 μg against A549, HeLa, HT29 and A375 cells, respectively. Followed by reference standard, Doxorubicin showed IC50 values of 17.53 ± 0.07 μg, 14.12 ± 0.30 μg, 17.44 ± 0.25 μg and 21.44 ± 0.40 μg against A549, HeLa, HT29 and A375 cells, respectively. AO/EtBr and DNA fragmentation assay confirmed the occurrence of apoptosis in the A549 cell line after the treatment of EAF. These results suggest that H. uninervis is a promising source of cytotoxicity against A549 cells and could be considered as a source for new lead structures to the development of new anticancer drugs.
Keywords:
Halodule uninervis; GC-MS; Anticancer activity; AO/EtBr staining; DNA fragmentationDOI
https://doi.org/10.25004/IJPSDR.2021.130611References
Mathers CD. History of global burden of disease assessment at the World Health Organization. Archives of Public Health. 2020; 78(1):1-13.
Akindele AJ, Wani ZA, Sharma S, Mahajan G, Satti NK, Adeyemi OO, Mondhe DM, Saxena AK. In vitro and in vivo anticancer activity of root extracts of Sansevieria liberica Gerome and Labroy (Agavaceae). Evidence-Based Complementary and Alternative Medicine. 2015; 2015.
Schwartsmann G, Ratain MJ, Cragg GM, Wong JE, Saijo N, Parkinson DR, Fujiwara Y, Pazdur R, Newman DJ, Dagher R, Di Leone L. Anticancer drug discovery and development throughout the world. Journal of clinical oncology. 2002; 20(18; SUPP): 47s-59s.
Pucci C, Martinelli C, Ciofani G. Innovative approaches for cancer treatment: Current perspectives and new challenges. Ecancermedicalscience. 2019; 13.
O’Reilly M, Mellotte G, Ryan B, O’Connor A. Gastrointestinal side effects of cancer treatments. Therapeutic Advances in Chronic Disease. 2020; 11:2040622320970354.
Gordaliza M. Natural products as leads to anticancer drugs. Clinical and Translational Oncology. 2007; 9(12):767-776.
Cragg GM, Pezzuto JM. Natural products as a vital source for the discovery of cancer chemotherapeutic and chemopreventive agents. Medical Principles and Practice. 2016; 25(Suppl. 2):41-59.
Martins A, Vieira H, Gaspar H, Santos S. Marketed marine natural products in the pharmaceutical and cosmeceutical industries: Tips for success. Marine drugs. 2014; 12(2):1066-1101.
Bibi F, Naseer MI, Hassan AM, Yasir M, Al-Ghamdi AAK, Azhar EI. Diversity and antagonistic potential of bacteria isolated from marine grass Halodule uninervis. 3 Biotech. 2018; 8(1):1-8.
Little RD, Nishiguchi GA. Synthetic efforts toward, and biological activity of, thyrsiferol and structurally-related analogues. In Studies in Natural Products Chemistry. 2008; (Vol. 35, pp. 3-56). Elsevier.
Victor SP, Sharma CP. Anti-inflammatory drug delivery systems using marine products. In Functional Marine Biomaterials. 2015; (pp. 137-147). Woodhead Publishing.
Dewsbury BM, Bhat M, Fourqurean JW. A review of seagrass economic valuations: gaps and progress in valuation approaches. Ecosystem Services. 2016; 18:68-77.
de la Torre-Castro M, Rönnbäck P. Links between humans and seagrasses—an example from tropical East Africa. Ocean & Coastal Management. 2004; 47(7-8):361-387.
Kontiza I, Vagias C, Jakupovic J, Moreau D, Roussakis C, Roussis V. Cymodienol and cymodiene: new cytotoxic diarylheptanoids from the sea grass Cymodocea nodosa. Tetrahedron letters. 2005; 46(16):2845-2847.
Styshova ON, Popov AM, Artyukov AA, Klimovich AA. Main constituents of polyphenol complex from seagrasses of the genus Zostera, their antidiabetic properties and mechanisms of action. Experimental and therapeutic medicine. 2017; 13(5):1651-1659.
Hua KF, Hsu HY, Su YC, Lin IF, Yang SS, Chen YM, Chao LK. Study on the antiinflammatory activity of methanol extract from seagrass Zostera japonica. Journal of agricultural and food chemistry. 2006; 54(2):306-311.
Ross C, Puglisi MP, Paul VJ. Antifungal defenses of seagrasses from the Indian River Lagoon, Florida. Aquatic Botany. 2008; 88(2):134-141.
Kontiza I, Stavri M, Zloh M, Vagias C, Gibbons S, Roussis V. New metabolites with antibacterial activity from the marine angiosperm Cymodocea nodosa. Tetrahedron. 2008; 64(8):1696-1702.
Kannan RRR, Arumugam R, Anantharaman P. Antibacterial potential of three seagrasses against human pathogens. Asian Pacific Journal of Tropical Medicine. 2010; 3(11):890-893.
Den Hartog C. The seagrasses of the world North Holland Publ. Co., Amsterdam, 1970, 275pp.
Kuo J, Den Hartog C. Seagrass taxonomy and identification key. Global seagrass research methods. 2001; 33:31-58.
Gumgumjee NM, Bukhari DA, Alshehri WA, Hajar AS. Antibacterial activity of Halodule uninervis leaves extracts against some bacterial pathogens strains. Pharmacophore. 2018; 9(2):52-59.
Karthikeyan R, Sundarapandian M. Antidiabetic activity of methanolic extract of Halodule uninervis in streptozotocin-induced diabetic mice. Journal of Pharmaceutical Sciences and Research. 2017; 9(10):1864-1868.
Harborne JB. Phytochemical Methods. A Guide to Modern Techniques of Plant Analysis, Chapman and Hall, London Ltd., 1973.
Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food chemistry. 1999; 64(4):555-559.
Chun OK, Kim DO, Lee CY. Superoxide radical scavenging activity of the major polyphenols in fresh plums. Journal of agricultural and food chemistry. 2003; 51(27):8067-8072.
Mosdam T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxic assay. J. Immunol. Methods. 1983; 65:55-63.
Martins A, Vieira H, Gaspar H, Santos S. Marketed marine natural products in the pharmaceutical and cosmeceutical industries: Tips for success. Marine drugs. 2014; 12(2):1066-1101.
Martins A, Vieira H, Gaspar H, Santos S. Marketed marine natural products in the pharmaceutical and cosmeceutical industries: Tips for success. Marine drugs. 2014; 12(2):1066-1101.
Baehaki A, Supriadi A, Pratama MC. Antioxidant activity of methanol extract of Halodule uninervis seagrass from the coastal of Lampung, Indonesia. Research Journal of Pharmaceutical, Biological and Chemical Sciences. 2016; 7(3):1173-1177.
Panche AN, Diwan AD, Chandra SR. Flavonoids: an overview. Journal of nutritional science. 2016; 5.
Kumar S, Pandey AK. Chemistry and biological activities of flavonoids: an overview. The scientific world journal. 2013; 2013.
Lee YK, Yuk DY, Lee JW, Lee SY, Ha TY, Oh KW, Yun YP, Hong JT. (−)-Epigallocatechin-3-gallate prevents lipopolysaccharide-induced elevation of beta-amyloid generation and memory deficiency. Brain research. 2009; 1250:164-174.
Rodríguez-García C, Sánchez-Quesada C, Gaforio JJ. Dietary flavonoids as cancer chemopreventive agents: An updated review of human studies. Antioxidants. 2019; 8(5):137.
Starlin T, Prabha PS, Thayakumar BKA, Gopalakrishnan VK. Screening and GC-MS profiling of ethanolic extract of Tylophora pauciflora. Bioinformation. 2019; 15(6):425.
Liu K, Liu PC, Liu R, Wu X. Dual AO/EB staining to detect apoptosis in osteosarcoma cells compared with flow cytometry. Medical science monitor basic research. 2015; 21:15.
Swanson SM, Ijaz A, Fahning ML. The use of acridine orange and ethidium bromide to determine the viability of pre-implantation mouse embryos cultured in vitro. British Veterinary Journal. 1987; 143(4):306-311.
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