Expression of Vascular Endothelial Growth Factor and Anti-Proliferative Activity of Flaxseed Oil Alone and In Combination with Mefenamic Acid in Cell Lines

Authors

  • Noor Alzubaidy Department of Pharmacology, College of Medicine, Al-Nahrain University, Baghdad, Iraq.
  • Hayder Sahib College of Pharmacy, Al-Nahrain University, Baghdad, Iraq.

DOI:

https://doi.org/10.31351/vol33iss1pp46-53

Keywords:

VEGF, MTT, KS, HUVEC, FO, COMB, RT- PCR

Abstract

The flaxseed oil contains alpha-linolenic acid, lignans and flavonoids. It possesses anti-inflammatory, antioxidant, and tumor-suppressive effects. The purpose of this research was to explore the anti-proliferative effect of flaxseed oil alone and flaxseed oil in combination with mefenamic acid on human umbilical vein endothelial cells and Kaposi sarcoma cells and to identify the vascular endothelial growth factor gene expression. The anti-proliferative effects of flaxseed oil alone and in combination with mefenamic acid were investigated in vitro by the 3-(4, 5-Dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay. The effects of flaxseed oil alone and its combination with mefenamic acid were tested on the expression of vascular endothelial growth factor genes using real-time polymerase chain reaction. The results showed that the IC50 of flaxseed oil for human umbilical vein endothelial cells was 29.72 µg/ml and for Kaposi sarcoma cells was 16.48 µg/ml; moreover, the IC50 for flaxseed oil in combination with mefenamic acid for human umbilical vein endothelial cells was 252 µg/ml and for Kaposi sarcoma cells was 147.6 µg/ml. These outcomes revealed that flaxseed oil alone and its combination with mefenamic acid possess anti-angiogenic activity in a concentration-dependent manner which supported by significant decline in vascular endothelial growth factor expression.

References

Rizvi QH, Sharma V, Shams R, Dar AH, Jan B, Manzoor A. Extraction of oil from flaxseed using three phase partitioning. Journal of Postharvest Technology. 2021;9:35-45.

Wrigley CW, Corke H, Seetharaman K, Faubion J, editors. Encyclopedia of food grains. Academic Press; 2015 Dec 17.

Tang ZX, Ying RF, Lv BF, Yang LH, Xu Z, Yan LQ, Bu JZ, Wei YS. Flaxseed oil: Extraction, Health benefits and products. Quality Assurance and Safety of Crops & Foods. 2021 Jan 2;13(1):1-9.

Yang R, Xue L, Zhang L, Wang X, Qi X, Jiang J, Yu L, Wang X, Zhang W, Zhang Q, Li P. Phytosterol contents of edible oils and their contributions to estimated phytosterol intake in the Chinese diet. Foods. 2019 Aug 9;8(8):334.

Singh A. Sitosterol as an antioxidant in frying oils. Food chemistry. 2013 Apr 15;137(1-4):62-7.

Jiang L, Zhao X, Xu J, Li C, Yu Y, Wang W, Zhu L. The protective effect of dietary phytosterols on cancer risk: a systematic meta-analysis. Journal of Oncology. 2019 Jun 23;2019.

Woyengo TA, Ramprasath VR, Jones PJ. Anticancer effects of phytosterols. European journal of clinical nutrition. 2009 Jul;63(7):813-20.

Deng Q, Yu X, Ma F, Xu J, Huang F, Huang Q, Sheng F. Comparative analysis of the in-vitro antioxidant activity and bioactive compounds of flaxseed in China according to variety and geographical origin. International journal of Food properties. 2017 Dec 31;20(sup3):S2708-22.

Li X, Li J, Dong S, Li Y, Wei L, Zhao C, Li J, Liu X, Wang Y. Effects of germination on tocopherol, secoisolarlciresinol diglucoside, cyanogenic glycosides and antioxidant activities in flaxseed (Linum usitatissimum L.). International Journal of Food Science & Technology. 2019 Jul;54(7):2346-54.

Abraham A, Kattoor AJ, Saldeen T, Mehta JL. Vitamin E and its anticancer effects. Critical reviews in food science and nutrition. 2019 Sep 25;59(17):2831-8.

Nechipurenko S, Vereshchagin N, Shilina Y, Efremov S, Moskvin Y, Umirbekova Z. The isolation of lignan containing fractions from flaxseed Linum usitatissimum L. International Journal of Biology and Chemistry. 2021 Jul 1;14(1):177-83.

Gokhale S, & Sahu AN. Pharmacological properties of flaxseed, Linum usitatissimum Linn., as a potential medicinal plant: An overview. World Journal of Pharmaceutical Sciences.2016.

Santos HO, Price JC, Bueno AA. Beyond fish oil supplementation: the effects of alternative plant sources of omega-3 polyunsaturated fatty acids upon lipid indexes and cardiometabolic biomarkers—an overview. Nutrients. 2020 Oct 16;12(10):3159.

Belmonte AM, Macchioni P, Minelli G, Scutaru C, Volpelli LA, Fiego DP. Effects of high linolenic acid diet supplemented with synthetic or natural antioxidant mix on live performance, carcass traits, meat quality and fatty acid composition of Longissimus thoracis et lumborum muscle of medium-heavy pigs. Italian Journal of Food Science. 2021 Jun 30;33(2):117-28.

Abdulazeem L, Tariq A, Jasim SA. An Investigation of Vascular Endothelial Growth Factor (VEGFR-1 and VEGFR-2) in burn wound healing. Archives of Razi Institute. 2022 Apr 30;77(2):747-51.

Park HJ, Zhang Y, Georgescu SP, Johnson KL, Kong D, Galper JB. Human umbilical vein endothelial cells and human dermal microvascular endothelial cells offer new insights into the relationship between lipid metabolism and angiogenesis. Stem cell reviews. 2006 Jun;2:93-101.

Wang Y, Liang Y, Vanhoutte PM. SIRT1 and AMPK in regulating mammalian senescence: a critical review and a working model. Federation of European Biochemical Societies (FEBS) letters. 2011 Apr 6;585(7):986-94.

Kocherova I, Bryja A, Mozdziak P, Angelova Volponi A, Dyszkiewicz-Konwińska M, Piotrowska-Kempisty H, Antosik P, Bukowska D, Bruska M, Iżycki D, Zabel M. Human umbilical vein endothelial cells (HUVECs) co-culture with osteogenic cells: from molecular communication to engineering prevascularised bone grafts. Journal of clinical medicine. 2019 Oct 3;8(10):1602.

Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of immunological methods. 1983 Dec 16;65(1-2):55-63.

Ponchel F, Toomes C, Bransfield K, Leong FT, Douglas SH, Field SL, Bell SM, Combaret V, Puisieux A, Mighell AJ, Robinson PA. Real-time PCR based on SYBR-Green I fluorescence: an alternative to the TaqMan assay for a relative quantification of gene rearrangements, gene amplifications and micro gene deletions. BMC biotechnology. 2003 Dec;3:1-3.

Kridel SJ, Axelrod F, Rozenkrantz N, Smith JW. Orlistat is a novel inhibitor of fatty acid synthase with antitumor activity. Cancer research. 2004 Mar 15;64(6):2070-5.

Tan ML, Sulaiman SF, Najimuddin N, Samian MR, Muhammad TT. Methanolic extract of Pereskia bleo (Kunth) DC.(Cactaceae) induces apoptosis in breast carcinoma, T47-D cell line. Journal of ethnopharmacology. 2005 Jan 4;96(1-2):287-94.

Shen Y, Chen G, Xiao A, Xie Y, Liu L, Cao Y. In vitro effect of flaxseed oil and α-linolenic acid against the toxicity of lipopolysaccharide (LPS) to human umbilical vein endothelial cells. Inflammopharmacology. 2018 Apr;26:645-54.

Sahib HB, Al-Zubaidy AA, Hussain SM, Jassim GA. The anti angiogenic activity of vitex agnus castus leaves extracts. International Journal of Pharmacy and Pharmaceutical Sciences. 2014;6(1):863-9.

Al-Zubaidy NA, Sahib HB. The Antiangiogenic Activity of Flaxseed Oil Alone and Combination with Mefenamic Acid in Vivo and in Vitro Assay. Asian Pacific Journal of Cancer Prevention. 2022 May 1;23(5):1711-7.

Ajith TA, Janardhanan KK. Cytotoxic and antitumor activities of a polypore macrofungus, Phellinus rimosus (Berk) Pilat. Journal of ethnopharmacology. 2003 Feb 1;84(2-3):157-62.

Yang J, Jadhav PD, Reaney MJ, Sammynaiken R, Yang J. A novel formulation significantly increases the cytotoxicity of flaxseed orbitides (linusorbs) LOB3 and LOB2 towards human breast cancer MDA-MB-231 cells. Die Pharmazie-An International Journal of Pharmaceutical Sciences. 2019 Sep 1;74(9):520-2.

Buckner A. Flaxseed oil induces apoptosis in the aggressive murine melanoma cell line B16-BL6.Doctoral dissertation, Laurentian University of Sudbury, 2014.‏

Buckner AL, Buckner CA, Montaut S, Lafrenie RM. Treatment with flaxseed oil induces apoptosis in cultured malignant cells. Heliyon. 2019 Aug 1;5(8):e02251.

Sousa Moreira I, Alexandrino Fernandes P, Joao Ramos M. Vascular endothelial growth factor (VEGF) inhibition-a critical review. Anti-cancer agents in medicinal chemistry (formerly current medicinal chemistry-anti-cancer agents). 2007 Mar 1;7(2):223-45.

Nash AD, Baca M, Wright C, Scotney PD. The biology of vascular endothelial growth factor-B (VEGF-B). Pulmonary pharmacology & therapeutics. 2006 Feb 1;19(1):61-9.

Niu G, Chen X. Vascular endothelial growth factor as an anti-angiogenic target for cancer therapy. Current drug targets. 2010 Aug 1;11(8):1000-17.

Zingg JM. Vitamin E: regulatory role on signal transduction. International Union of Biochemistry and Molecular Biology (IUBMB) life. 2019 Apr;71(4):456-78.

Mason JK, Thompson LU. Flaxseed and its lignan and oil components: can they play a role in reducing the risk of and improving the treatment of breast cancer?. Applied Physiology, Nutrition, and Metabolism. 2014;39(6):663-78.

Kangsamaksin T, Chaithongyot S, Wootthichairangsan C, Hanchaina R, Tangshewinsirikul C, Svasti J. Lupeol and stigmasterol suppress tumor angiogenesis and inhibit cholangiocarcinoma growth in mice via downregulation of tumor necrosis factor-α. The Public Library of Science (PloS one). 2017 Dec 12;12(12):e0189628.

Imamura T, Nguyen A, Rodgers D, Kim G, Raikhelkar J, Sarswat N, Kalantari S, Smith B, Chung B, Narang N, Juricek C. Omega-3 therapy is associated with reduced gastrointestinal bleeding in patients with continuous-flow left ventricular assist device. Circulation: Heart Failure. 2018 Oct;11(10):e005082.

Scoditti E, Calabriso N, Massaro M, Pellegrino M, Storelli C, Martines G, De Caterina R, Carluccio MA. Mediterranean diet polyphenols reduce inflammatory angiogenesis through MMP-9 and COX-2 inhibition in human vascular endothelial cells: a potentially protective mechanism in atherosclerotic vascular disease and cancer. Archives of biochemistry and biophysics. 2012 Nov 15;527(2):81-9.

Ezzat SM, Shouman SA, Elkhoely A, Attia YM, Elsesy MS, El Senousy AS, Choucry MA, El Gayed SH, El Sayed AA, Sattar EA, El Tanbouly N. Anticancer potentiality of lignan rich fraction of six Flaxseed cultivars. Scientific Reports. 2018 Jan 11;8(1):1-6.

Tarnawski AS, Jones MK. Inhibition of angiogenesis by NSAIDs: molecular mechanisms and clinical implications. Journal of molecular medicine. 2003 Oct;81:627-36.

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Published

2024-03-26

How to Cite

1.
Alzubaidy N, Sahib H. Expression of Vascular Endothelial Growth Factor and Anti-Proliferative Activity of Flaxseed Oil Alone and In Combination with Mefenamic Acid in Cell Lines. IJPS [Internet]. 2024 Mar. 26 [cited 2024 Oct. 9];33(1):46-53. Available from: https://bijps.uobaghdad.edu.iq/index.php/bijps/article/view/2261