The evaluation of antioxidant and anticancer effects of Lepidium Sativum Subsp Spinescens L. methanol extract on cancer cells

Sahabettin Selek; Ismail Koyuncu; Hifa Gulru Caglar; Ibrahim Bektas; Mustafa Abdullah Yilmaz; Ataman Gonel; Enes Akyuz

doi:10.14715/cmb/2018.64.3.12

Issue

Vol. 64 No. 3: Issue 3

Issue Published : March 3, 2018

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The evaluation of antioxidant and anticancer effects of Lepidium Sativum Subsp Spinescens L. methanol extract on cancer cells

https://doi.org/10.14715/cmb/2018.64.3.12

Sahabettin Selek

Department of Medical Biochemistry, Faculty of Medicine, Bezmialem Vakif University, Istanbul, Turkey

http://orcid.org/0000-0003-1235-3957

Ismail Koyuncu

Department of Biochemistry, Faculty of Medicine, Harran University, Sanliurfa, Turkey

Hifa Gulru Caglar

Department of Medical Biochemistry, Faculty of Medicine, Bezmialem Vakif University, Istanbul, Turkey

Ibrahim Bektas

Department of Pharmacy Services, Health Services Vocational School, Harran University, Sanliurfa, Turkey

Mustafa Abdullah Yilmaz

Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Dicle University, Diyarbakir, Turkey

Ataman Gonel

Department of Biochemistry, Faculty of Medicine, Harran University, Sanliurfa, Turkey

Enes Akyuz

Department of Biophysics, Faculty of Medicine, Bezmialem Vakif University, Istanbul, Turkey

Corresponding Author(s) : Sahabettin Selek

sahselek@gmail.com

Cellular and Molecular Biology, Vol. 64 No. 3: Issue 3
Article Published : February 28, 2018

Abstract

In recent years, there is an increased research interest for plants which are natural sources of antioxidants. Lepidium sativum Subsp spinescens L., commonly found in South West Asia, is a plant known as a healthy nutritional source containing bio-molecules that carry anti-hypertensive, hypoglycemic, anti-asthmatic, antispasmodic, hepato-protective, chemoprotective, anti-inflammatory and anti-oxidant effects. In this study, we aimed to investigate the antioxidant content and activity of Lepidium sativum Subsp spinescens L. methanol extract on cancer cells. Methanol extract of dried Lepidium sativum Subsp spinescens L. was prepared. Total amount of phenolic compounds was determined by Slinkard and Singleton method using Folin-Ciocalteu reagent. Total flavonoid amount was determined according to Zhishen method. Antioxidant activity of the extract was evaluated by CUPRAC and ABTS radical scavenging activity assays. Cytotoxic effects of the plant extract on colon and endometrium cancer cells, and human peripheral lymphocyte cells were investigated in vitro by MTT and neutral red assays. Furthermore, the plant extract was investigated for necrotic effects by LDH assay; apoptotic activity by DNA ladder fragmentation, ELISA and acridine orange/ethidium bromide staining; and genotoxic effect by comet assay methods. Methanol extract of Lepidium sativum Subsp spinescens L. was found to have a high content of phenolic and flavonoid compounds. The extract showed significant antioxidant activity and also cytotoxic activity on colon and endometrium cancer cells in a concentration-dependent manner. Apoptotic activity and genotoxic effects were significantly increased, especially with 200 Î¼g/ml concentrations at 48 hours incubation. In conclusion, it was determined that the extract evaluated in this study could be a natural source of antioxidants. Further molecular studies explaining chemo-preventive and chemotherapeutic effects on cancer cells are required to support anticancer efficacy of the plant.

Keywords

Lepidium sativum Subsp spinescens L. Antioxidant activity Anticancer activity.

Selek, S., Koyuncu, I., Caglar, H. G., Bektas, I., Yilmaz, M. A., Gonel, A., & Akyuz, E. (2018). The evaluation of antioxidant and anticancer effects of Lepidium Sativum Subsp Spinescens L. methanol extract on cancer cells. Cellular and Molecular Biology, 64(3), 72–80. https://doi.org/10.14715/cmb/2018.64.3.12

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References

Global Burden of Disease Cancer C, Fitzmaurice C, Allen C, Barber RM, Barregard L, Bhutta ZA, et al. Global, Regional, and National Cancer Incidence, Mortality, Years of Life Lost, Years Lived With Disability, and Disability-Adjusted Life-years for 32 Cancer Groups, 1990 to 2015: A Systematic Analysis for the Global Burden of Disease Study. JAMA oncology. 2017;3(4):524-48.
Pisoschi AM, Pop A. The role of antioxidants in the chemistry of oxidative stress: A review. Eur J Med Chem. 2015;97:55-74.
Swanson H. Flavonoids, Inflammation and Cancer: World Scientific; 2015.
Poljsak B, Suput D, Milisav I. Achieving the balance between ROS and antioxidants: when to use the synthetic antioxidants. Oxid Med Cell Longev. 2013;2013:956792.
Reuter S, Gupta SC, Chaturvedi MM, Aggarwal BB. Oxidative stress, inflammation, and cancer: how are they linked? Free Radic Biol Med. 2010;49(11):1603-16.
Hwang YJ, Lee EJ, Kim HR, Hwang KA. In vitro antioxidant and anticancer effects of solvent fractions from Prunella vulgaris var. lilacina. BMC Complement Altern Med. 2013;13:310.
Gokavi SS, Malleshi NG, Guo M. Chemical composition of garden cress (Lepidium sativum) seeds and its fractions and use of bran as a functional ingredient. Plant Foods Hum Nutr. 2004;59(3):105-11.
Diwakar BT, Dutta PK, Lokesh BR, Naidu KA. Physicochemical Properties of Garden Cress (Lepidium sativum L.) Seed Oil. Journal of the American Oil Chemists' Society. 2009;87(5):539-48.
Kassie F, Rabot S, Uhl M, Huber W, Qin HM, Helma C, et al. Chemoprotective effects of garden cress (Lepidium sativum) and its constituents towards 2-amino-3-methyl-imidazo[4,5-f]quinoline (IQ)-induced genotoxic effects and colonic preneoplastic lesions. Carcinogenesis. 2002;23(7):1155-61.
Pramod Jyotiram Kasabe PNP, Dayanand D Kamble, Padma Dandge. Nutritional, elemental analysis and antioxidant activity of garden cress (Lepidium sativum L.) seeds. Int J Pharm Pharm Sci 43 2012:392-5.
Doke Snehal MG. Garden cress (Lepidium sativum L.) seed-An important medicinal source. J Nat Prod Plant Resour 41 2014:69-80.
Diwakar BT, Dutta PK, Lokesh BR, Naidu KA. Bio-availability and metabolism of n-3 fatty acid rich garden cress (Lepidium sativum) seed oil in albino rats. Prostaglandins Leukot Essent Fatty Acids. 2008;78(2):123-30.
Singleton VL, Orthofer R, Lamuela-Raventós RM. [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods in enzymology. 1999;299:152-78.
Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem. 1999;64(4):555-9.
Apak R, Güçlü K, í–zyürek M, Karademir SE. Novel total antioxidant capacity index for dietary polyphenols and vitamins C and E, using their cupric ion reducing capability in the presence of neocuproine: CUPRAC method. Journal of agricultural and food chemistry. 2004;52(26):7970-81.
Repetto G, del Peso A, Zurita JL. Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nat Protoc. 2008;3(7):1125-31.
Vermes I, Haanen C, Steffens-Nakken H, Reutellingsperger C. A novel assay for apoptosis flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin V. Journal of immunological methods. 1995;184(1):39-51.
Tice R, Andrews P, Singh N. The single cell gel assay: a sensitive technique for evaluating intercellular differences in DNA damage and repair. DNA damage and repair in human tissues: Springer; 1990. p. 291-301.
Isik M, Beydemir S, Yilmaz A, Naldan ME, Aslan HE, Gulcin I. Oxidative stress and mRNA expression of acetylcholinesterase in the leukocytes of ischemic patients. Biomed Pharmacother. 2017;87:561-7.
Gulcin I, Beydemir S. Phenolic compounds as antioxidants: carbonic anhydrase isoenzymes inhibitors. Mini Rev Med Chem. 2013;13(3):408-30.
Tohma H, Isik M, Korkmaz M, Bursal E, Gulcin I, Koksal E. Determination of Antioxidant Properties of Gypsophila bitlisensis Bark. Int J Pharmacol. 2015;11(4):366-71.
Gülçin ć°. Antioxidant properties of resveratrol: A structure–activity insight. Innov Food Sci Emerg. 2010;11(1):210-8.
Carocho M, Ferreira IC. A review on antioxidants, prooxidants and related controversy: natural and synthetic compounds, screening and analysis methodologies and future perspectives. Food Chem Toxicol. 2013;51:15-25.
Balasundram N, Sundram K, Samman S. Phenolic compounds in plants and agri-industrial by-products: Antioxidant activity, occurrence, and potential uses. Food Chem. 2006;99(1):191-203.
Zia-Ul-Haq M, Ahmad S, Calani L, Mazzeo T, Del Rio D, Pellegrini N, et al. Compositional study and antioxidant potential of Ipomoea hederacea Jacq. and Lepidium sativum L. seeds. Molecules. 2012;17(9):10306-21.
Athar M, Back JH, Tang X, Kim KH, Kopelovich L, Bickers DR, et al. Resveratrol: a review of preclinical studies for human cancer prevention. Toxicol Appl Pharmacol. 2007;224(3):274-83.
Zhong Y, Chiou Y-S, Pan M-H, Ho C-T, Shahidi F. Protective effects of epigallocatechin gallate (EGCG) derivatives on azoxymethane-induced colonic carcinogenesis in mice. J Funct Foods. 2012;4(1):323-30.
Siah SD, Konczak I, Agboola S, Wood JA, Blanchard CL. In vitro investigations of the potential health benefits of Australian-grown faba beans (Vicia faba L.): chemopreventative capacity and inhibitory effects on the angiotensin-converting enzyme, alpha-glucosidase and lipase. Br J Nutr. 2012;108 Suppl 1:S123-34.
AlObaidi LAH. Study the Anticancer Effect of Lepidium sativum Leaves Extract on Squamous Cell Carcinoma (CAL-27) Cell Lines. Journal of Natural Sciences Research 2014 4(17).
Mahassni SH, Al-Reemi RM. Apoptosis and necrosis of human breast cancer cells by an aqueous extract of garden cress (Lepidium sativum) seeds. Saudi J Biol Sci. 2013;20(2):131-9.

References

Global Burden of Disease Cancer C, Fitzmaurice C, Allen C, Barber RM, Barregard L, Bhutta ZA, et al. Global, Regional, and National Cancer Incidence, Mortality, Years of Life Lost, Years Lived With Disability, and Disability-Adjusted Life-years for 32 Cancer Groups, 1990 to 2015: A Systematic Analysis for the Global Burden of Disease Study. JAMA oncology. 2017;3(4):524-48.

Pisoschi AM, Pop A. The role of antioxidants in the chemistry of oxidative stress: A review. Eur J Med Chem. 2015;97:55-74.

Swanson H. Flavonoids, Inflammation and Cancer: World Scientific; 2015.

Poljsak B, Suput D, Milisav I. Achieving the balance between ROS and antioxidants: when to use the synthetic antioxidants. Oxid Med Cell Longev. 2013;2013:956792.

Reuter S, Gupta SC, Chaturvedi MM, Aggarwal BB. Oxidative stress, inflammation, and cancer: how are they linked? Free Radic Biol Med. 2010;49(11):1603-16.

Hwang YJ, Lee EJ, Kim HR, Hwang KA. In vitro antioxidant and anticancer effects of solvent fractions from Prunella vulgaris var. lilacina. BMC Complement Altern Med. 2013;13:310.

Gokavi SS, Malleshi NG, Guo M. Chemical composition of garden cress (Lepidium sativum) seeds and its fractions and use of bran as a functional ingredient. Plant Foods Hum Nutr. 2004;59(3):105-11.

Diwakar BT, Dutta PK, Lokesh BR, Naidu KA. Physicochemical Properties of Garden Cress (Lepidium sativum L.) Seed Oil. Journal of the American Oil Chemists' Society. 2009;87(5):539-48.

Kassie F, Rabot S, Uhl M, Huber W, Qin HM, Helma C, et al. Chemoprotective effects of garden cress (Lepidium sativum) and its constituents towards 2-amino-3-methyl-imidazo[4,5-f]quinoline (IQ)-induced genotoxic effects and colonic preneoplastic lesions. Carcinogenesis. 2002;23(7):1155-61.

Pramod Jyotiram Kasabe PNP, Dayanand D Kamble, Padma Dandge. Nutritional, elemental analysis and antioxidant activity of garden cress (Lepidium sativum L.) seeds. Int J Pharm Pharm Sci 43 2012:392-5.

Doke Snehal MG. Garden cress (Lepidium sativum L.) seed-An important medicinal source. J Nat Prod Plant Resour 41 2014:69-80.

Diwakar BT, Dutta PK, Lokesh BR, Naidu KA. Bio-availability and metabolism of n-3 fatty acid rich garden cress (Lepidium sativum) seed oil in albino rats. Prostaglandins Leukot Essent Fatty Acids. 2008;78(2):123-30.

Singleton VL, Orthofer R, Lamuela-Raventós RM. [14] Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Methods in enzymology. 1999;299:152-78.

Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem. 1999;64(4):555-9.

Apak R, Güçlü K, í–zyürek M, Karademir SE. Novel total antioxidant capacity index for dietary polyphenols and vitamins C and E, using their cupric ion reducing capability in the presence of neocuproine: CUPRAC method. Journal of agricultural and food chemistry. 2004;52(26):7970-81.

Repetto G, del Peso A, Zurita JL. Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nat Protoc. 2008;3(7):1125-31.

Vermes I, Haanen C, Steffens-Nakken H, Reutellingsperger C. A novel assay for apoptosis flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin V. Journal of immunological methods. 1995;184(1):39-51.

Tice R, Andrews P, Singh N. The single cell gel assay: a sensitive technique for evaluating intercellular differences in DNA damage and repair. DNA damage and repair in human tissues: Springer; 1990. p. 291-301.

Isik M, Beydemir S, Yilmaz A, Naldan ME, Aslan HE, Gulcin I. Oxidative stress and mRNA expression of acetylcholinesterase in the leukocytes of ischemic patients. Biomed Pharmacother. 2017;87:561-7.

Gulcin I, Beydemir S. Phenolic compounds as antioxidants: carbonic anhydrase isoenzymes inhibitors. Mini Rev Med Chem. 2013;13(3):408-30.

Tohma H, Isik M, Korkmaz M, Bursal E, Gulcin I, Koksal E. Determination of Antioxidant Properties of Gypsophila bitlisensis Bark. Int J Pharmacol. 2015;11(4):366-71.

Gülçin ć°. Antioxidant properties of resveratrol: A structure–activity insight. Innov Food Sci Emerg. 2010;11(1):210-8.

Carocho M, Ferreira IC. A review on antioxidants, prooxidants and related controversy: natural and synthetic compounds, screening and analysis methodologies and future perspectives. Food Chem Toxicol. 2013;51:15-25.

Balasundram N, Sundram K, Samman S. Phenolic compounds in plants and agri-industrial by-products: Antioxidant activity, occurrence, and potential uses. Food Chem. 2006;99(1):191-203.

Zia-Ul-Haq M, Ahmad S, Calani L, Mazzeo T, Del Rio D, Pellegrini N, et al. Compositional study and antioxidant potential of Ipomoea hederacea Jacq. and Lepidium sativum L. seeds. Molecules. 2012;17(9):10306-21.

Athar M, Back JH, Tang X, Kim KH, Kopelovich L, Bickers DR, et al. Resveratrol: a review of preclinical studies for human cancer prevention. Toxicol Appl Pharmacol. 2007;224(3):274-83.

Zhong Y, Chiou Y-S, Pan M-H, Ho C-T, Shahidi F. Protective effects of epigallocatechin gallate (EGCG) derivatives on azoxymethane-induced colonic carcinogenesis in mice. J Funct Foods. 2012;4(1):323-30.

Siah SD, Konczak I, Agboola S, Wood JA, Blanchard CL. In vitro investigations of the potential health benefits of Australian-grown faba beans (Vicia faba L.): chemopreventative capacity and inhibitory effects on the angiotensin-converting enzyme, alpha-glucosidase and lipase. Br J Nutr. 2012;108 Suppl 1:S123-34.

AlObaidi LAH. Study the Anticancer Effect of Lepidium sativum Leaves Extract on Squamous Cell Carcinoma (CAL-27) Cell Lines. Journal of Natural Sciences Research 2014 4(17).

Mahassni SH, Al-Reemi RM. Apoptosis and necrosis of human breast cancer cells by an aqueous extract of garden cress (Lepidium sativum) seeds. Saudi J Biol Sci. 2013;20(2):131-9.

Author biographies is not available.