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Anti-Proliferative Properties of Terminalia Sericea Burch. Ex Dc Leaf Extracts Against Caco2 and Hela Cancer Cell Lines

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Anti-Proliferative Properties of Terminalia Sericea Burch. Ex Dc Leaf Extracts Against Caco2 and Hela Cancer Cell Lines Pharmacogn J. 2018; 10(3):408-415. A Multifaceted Journal in the field of Natural Products and Pharmacognosy Original Article www.phcogj.com | www.journalonweb.com/pj | www.phcog.net Anti-Proliferative Properties of Terminalia sericea Burch. Ex Dc Leaf Extracts Against Caco2 and HeLa Cancer Cell Lines BiYun Gu1,2, Joseph Shalom1,3, Ian E. Cock1,3* ABSTRACT Introduction: Terminalia spp. are characterised by their high levels of antioxidant phytochemicals and several species have anticancer activity. This study examines the anti-proliferative activity of T. sericea leaf extracts against Caco2 and HeLa carcinoma cell proliferation. Methods: Solvent extracts were prepared from T. sericea leaves and their antioxidant capacities were determined by the DPPH free radical scavenging assay. Anti-proliferative activities against Caco2 and HeLa cancer cells were determined by an MTS based cell proliferation assay. Toxicity was determined using the Artemia franciscana nauplii bioassay. Results: The methanolic and aqueous T. sericea leaf extracts displayed high antioxidant capacities (equivalent to 150 and BiYun Gu1,2, 340 mg of ascorbic acid per gram of plant material extracted respectively). In contrast, the Joseph Shalom1,3, ethyl acetate, chloroform and hexane extracts had relatively low antioxidant contents (≤5 mg Ian E. Cock1,3* of ascorbic acid equivalents per gram of plant material extracted). The antioxidant contents of the T. sericea leaf extracts correlated with the ability of the extracts to inhibit proliferation 1School of Natural Sciences, of Caco2 and HeLa cancer cell lines. The high antioxidant methanolic and aqueous extracts were potent inhibitors of cell proliferation, with IC values 120-1400 µg/mL. The aqueous Nathan Campus, Griffith University, 50 170 Kessels Rd, Nathan, Queensland T. sericea leaf extract was particularly effective, with IC50 values of 528 and 120 µg/mL against 4111, AUSTRALIA Caco2 and HeLa cells respectively. The methanolic extract also displayed good, albeit 2 School of Pharmacy, substantially less potent, antiproliferative activity against HeLa cells, with an IC50 of 1358 µg/mL. Nanjing University of Chinese Medicine, In contrast, the lower antioxidant content extracts generally did not inhibit cancer cell prolif- Nanjing, CHINA. eration. Cell imaging studies detected morphological features consistent with apoptosis in 3Environmental Futures Research Caco2 cells exposed to sub-lethal concentrations of the methanolic and aqueous T. sericea Institute, Nathan Campus, Griffith leaf extracts, indicating that these extracts are functioning by cytotoxic mechanisms. The University, 170 Kessels Rd, Nathan, aqueous T. sericea leaf extract displayed low to moderate toxicity in the Artemia franciscana Queensland 4111, AUSTRALIA bioassay, with an LC50 value of 737 µg/mL. All other extracts were nontoxic. Conclusion: The Correspondence antiproliferative activity and low toxicity of the T. sericea methanolic and aqueous leaf extracts extracts against HeLa and Caco2 cancer cell lines indicates their potential in the treatment Ian E. Cock and prevention of some cancers. School of Natural Sciences, Key words: Combretaceae, Silver Cluster Leaf, Anticancer Activity, Antiproliferative Activity, Nathan Campus, Griffith University, Antioxidant Capacity, Apoptosis, DPPH. 170 Kessels Rd, Nathan, Queensland 4111, AUSTRALIA & Environmental Futures Research Institute, Nathan Campus, Griffith University, 170 Kessels Rd, INTRODUCTION Nathan, Queensland 4111, AUSTRALIA Phone no : 61-7-37357637 Epidemiological studies have shown that a diet rich in The relationship between cellular redox state and E-mail: [email protected] antioxidants is associated with a decreased incidence cancer progression has been particularly well studied. 1 History of chronic diseases. High antioxidant levels have also The induction of cellular oxidative stress has been 8,9 • Submission Date: 07-08-2017; been shown to act as a preventative against the devel­ linked with several types of cancer. Thus it is • Review completed: 31-08-2017; opment of degenerative diseases such as cancer,2 cardio­ possible that high antioxidant contents of many plants • Accepted Date: 02-11-2017 vascular diseases,3 neural degeneration,4 diabetes and may inhibit cancer formation and/or progression. 5 Studies into the antioxidant/pro­oxidant effects DOI : 10.5530/pj.2018.3.67 obesity. Phenolic phytochemicals are generally strong antioxidants.6 Their primary action involves the protection of extracts from various plant species have dem­ Article Available online onstrated that the ability of a plant extract to exert of cell constituents against oxidative damage through http://www.phcogj.com/v10/i3 antioxidant activity depends on multiple factors. the scavenging of free radicals, thereby averting their Aloe vera antioxidant components for example may Copyright deleterious effects on nucleic acids, proteins, and lipids function as either antioxidants or a pro­oxidants, © 2018 Phcog.Net. This is an open- in cells.6 Phenolics interact directly with receptors or access article distributed under the terms with their action being dependent upon their con­ 7 of the Creative Commons Attribution 4.0 enzymes involved in signal transduction, indicating centration.10 The Aloe vera anthraquinone aloe International license. that they play specific roles in human physiology via emodin exerts antioxidant behaviour at lower modulation of the cellular redox state. Common plant concentrations, yet acts as a pro­oxidant at high phenolic compounds include flavonoids, tannins, concentrations. In contrast, a different Aloe vera anthocyanins, and gallic acid. anthraquinone (aloin) has an antioxidant effect at Cite this article: Gu B, Shalom J, Cock IE. Anti-Proliferative Properties of Terminalia Sericea Burch. Ex Dc Leaf Extracts Against Caco2 and Hela Cancer Cell Lines. Pharmacog J. 2018;10(2):408- 15. 408 Pharmacognosy Journal, Vol 10, Issue 3, May-Jun, 2018 BiYun et al.: T. sericea extracts inhibit cancer cell growth higher concentrations, yet a pro­oxidant effect at low concentrations. South Africa and were sourced from the Walter Sisulu Botanical Gardens Thus, Aloe vera extracts and components may act as either antioxidants in Johannesburg, South Africa. The plant materials were identified by or as pro-oxidants, dependent on differing levels of the various constitu- Andrew Hankey, chief botanist at the Walter Sisulu Botanical Gardens ents, and on their ratios. Thus, although many plant species have very in Johannesburg, South Africa. Voucher specimens are stored in the high antioxidant contents, it is possible that the individual components School of Natural Sciences, Griffith University, Australia. All plant may act as either antioxidants or as pro-oxidants and thus may also be material was thoroughly air dried and ground into a coarse powder. The effective in the treatment of cancer, as well as in its prevention at differ- dried plant material was stored at ­30oC until use. Individual 1 g quantities ent concentrations. of the powdered T. sericea were weighed into separate tubes and 50 mL Similar pro-oxidant effects have been reported for other antioxidant of methanol, deionised water, ethyl acetate, chloroform or hexane were phytochemicals including flavonoids11 and tannins.12 Furthermore, the added. All solvents were obtained from Ajax Chemicals, Australia and presence of transition metal ions such as copper or iron in an extract were AR grade. The groundT. sericea leaf material was extracted in each o can further enhance the conversion of the antioxidant to the pro­oxidant solvent for 24h at 4 C with gentle shaking. The extracts were filtered state.13,14 The pro­oxidant/antioxidant effect of plant extracts is due to through filter paper (Whatman No. 54) under vacuum. The solvent a balance between the free radical scavenging activities and reducing extracts were dried in a vacuum oven and the aqueous extract was dried power of their phytochemical components. This can be explained using by freeze drying. The resultant dry extract was weighed and redissolved the antioxidant vitamin ascorbic acid as an example. Although ascorbic in 10 mL deionised water (containing 1 % DMSO). acid has well characterised antioxidant bioactivities, it acts as a pro­oxidant Qualitative phytochemical studies at high concentrations.15 This is due to the greater reducing power of ascorbic acid compared to its free radical scavenging activity. In the Phytochemical analysis of the extracts for the presence of saponins, presence of transition metal ions, ascorbic acid will function as a reducing phenolic compounds, flavonoids, polysteroids, triterpenoids, cardiac agent, reducing the metal ions. In this process, it is converted to a glycosides, anthraquinones, tannins and alkaloids was conducted by 27,28 pro­oxidant. Therefore, high dietary intake of ascorbic acid (or other previously described assays. antioxidants) in individuals with high iron levels (e.g. premature infants) Antioxidant capacity may result in unexpected health effects due to the induction of oxidative The antioxidant capacity of each sample was assessed using the DPPH damage to susceptible biomolecules.16,17,18 free radical scavenging method29 with modifications. Briefly, DPPH Plants of the genus Terminalia (family Combretaceae) are characterised 19 solution was prepared fresh each day as a 400 µM solution by dissolving by their high antioxidant contents. It has previously been postulated DPPH (Sigma) in AR grade methanol (Ajax, Australia). The initial
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