The Effect of Protein Extract of Licorice Root in Proliferation of HT-29 and CT26 Cancer Cell Lines

Document Type : Original Article (s)

Authors

1 Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan AND Immunology Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran

2 Associate Professor, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan

3 Assistant Professor, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan

4 Immunology Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran

5 PhD student, Immunology Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran

6 Assistant Professor, Immunology Asthma and Allergy Research Institute, Tehran University of Medical Sciences, Tehran, Iran

Abstract

Background: Gastrointestinal cancers, especially colon cancer, are of the most common causes of death in western countries. Already, herbal and complementary medicine has been considered as a supplement treatment. The licorice is an ancient plant in herbal medicine that has many health benefits such as anti-cancer and anti-inflammatory effects. So, the present study was designed to assess the effect of protein extract of licorice root on human colon cancer cell line (HT-29), murine colon cancer (CT26) and normal cells (HEK293) for considering the cell growth inhibition potential after treatment with the extracts.Methods: Protein extracts of licorice root powder was prepared after grinding in phosphate buffered saline (PBS) at room temperature. After the dialysis in buffer, the protein concentration was determined via Bradford method. The HT-29, CT-26 and HEK-293 cell lines were maintained in Dulbecco's Modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum. After 24 hour of incubation at 37°C, the cells were treated with the concentrations of 25, 50 and 100 µg/ml of licorice extract. Cytotoxicity was evaluated via MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. In addition, the inductions of apoptosis (Annecin-V-Fluos staining method) in the treated cells were evaluated using flow cytometric analysis.Findings: Up to 100 µg/ml of the protein extract had not toxin effect on normal cell proliferation (HEK293). However, the viability of CT26, HT29 and HEK293 was reduced by 29.3%, 42.5% and 70%, respectively. Moreover, the treated CT26, HT29 and HEK293 cells showed the apoptosis percentages of 47.72 ± 8.00 (P = 0.026), 34.93 ± 8.21 (P = 0.056) and 17.5 ± 6.5 (P = 0.07), respectively, in comparison with not-treated cells.Conclusion: It appears that protein extract of licorice root could inhibit the colon cancer cell line proliferation and can be used as an adjuvant treatment.

Keywords

References

  1. Haggar FA, Boushey RP. Colorectal cancer epidemiology: incidence, mortality, survival, and risk factors. Clin Colon Rectal Surg 2009; 22(4): 191-7.
  2. Khan R, Khan AQ, Lateef A, Rehman MU, Tahir M, Ali F, et al. Glycyrrhizic Acid Suppresses the Development of Precancerous Lesions via Regulating the Hyperproliferation, Inflammation, Angiogenesis and Apoptosis in the Colon of Wistar Rats. PLoS ONE 2013; 8(2): e56020.
  3. Tai CJ, Wang WC, Wang CK, Wu CH, Yang MD, Chang YJ, et al. Fermented Wheat Germ Extract Induced Cell Death and Enhanced Cytotoxicity of Cisplatin and 5-Fluorouracil on Human Hepatocellular Carcinoma Cells. Evid Based Complement Alternat Med 2013; 2013: 121725.
  4. Turpie AG, Thomson TJ. Carbenoxolone sodium in the treatment of gastric ulcer with special reference to side-effects. Gut 1965; 6(6): 591-4.
  5. Wang GS, Han ZW. The protective action of glycyrrhiza flavonoids against carbon tetrachloride hepatotoxicity in mice. Yao Xue Xue Bao 1993; 28(8): 572-6.
  6. Kroes BH, Beukelman CJ, van den Berg AJ, Wolbink GJ, van DH, Labadie RP. Inhibition of human complement by beta-glycyrrhetinic acid. Immunology 1997; 90(1): 115-20.
  7. Moon A, Kim SH. Effect of Glycyrrhiza glabra roots and glycyrrhizin on the glucuronidation in rats. Planta Med 1997; 63(2): 115-9.
  8. Pompei R, Flore O, Marccialis MA, Pani A, Loddo B. Glycyrrhizic acid inhibits virus growth and inactivates virus particles. Nature 1979; 281(5733): 689-90.
  9. Pompei R, Paghi L, Ingianni A, Uccheddu P. Glycyrrhizic acid inhibits influenza virus growth in embryonated eggs. Microbiologica 1983; 6(3): 247-50.
  10. Shimizu N, Tomoda M, Takada K, Gonda R. The core structure and immunological activities of glycyrrhizan UA, the main polysaccharide from the root of Glycyrrhiza uralensis. Chem Pharm Bull (Tokyo) 1992; 40(8): 2125-8.
  11. Fukai T, Marumo A, Kaitou K, Kanda T, Terada S, Nomura T. Anti-Helicobacter pylori flavonoids from licorice extract. Life Sci 2002; 71(12): 1449-63.
  12. Mir Heidar H. Herbal sciences. Tehran, Iran: Nashr Farhang Publication; 1994. [In Persian].
  13. Rafi MM, Vastano BC, Zhu N, Ho CT, Ghai G, Rosen RT, et al. Novel polyphenol molecule isolated from licorice root (Glycrrhiza glabra) induces apoptosis, G2/M cell cycle arrest, and Bcl-2 phosphorylation in tumor cell lines. J Agric Food Chem 2002; 50(4): 677-84.
  14. Jo EH, Kim SH, Ra JC, Kim SR, Cho SD, Jung JW, et al. Chemopreventive properties of the ethanol extract of chinese licorice (Glycyrrhiza uralensis) root: induction of apoptosis and G1 cell cycle arrest in MCF-7 human breast cancer cells. Cancer Lett 2005; 230(2): 239-47.
  15. Jung JI, Lim SS, Choi HJ, Cho HJ, Shin HK, Kim EJ, et al. Isoliquiritigenin induces apoptosis by depolarizing mitochondrial membranes in prostate cancer cells. J Nutr Biochem 2006; 17(10): 689-96.
  16. Wang ZY, Nixon DW. Licorice and cancer. Nutr Cancer 2001; 39(1): 1-11.
  17. Kim A, Yim NH, Yeul Ma J. Samsoeum, a traditional herbal medicine, elicits apoptotic and autophagic cell death by inhibiting Akt/mTOR and activating the JNK pathway in cancer cells. BMC Complementary and Alternative Medicine 2013; 13: 233.
  18. Su CC, Chen GW, Tan TW, Lin JG, Chung JG. Crude extract of garlic induced caspase-3 gene expression leading to apoptosis in human colon cancer cells. In Vivo 2006; 20(1): 85-90.
  19. Bouallagui Z, Han J, Isoda H, Sayadi S. Hydroxytyrosol rich extract from olive leaves modulates cell cycle progression in MCF-7 human breast cancer cells. Food Chem Toxicol 2011; 49(1): 179-84.
  20. Rossi T, Benassi L, Magnoni C, Ruberto AI, Coppi A, Baggio G. Effects of glycyrrhizin on UVB-irradiated melanoma cells. In Vivo 2005; 19(1): 319-22.
  21. Hsu YL, Kuo PL, Lin LT, Lin CC. Isoliquiritigenin inhibits cell proliferation and induces apoptosis in human hepatoma cells. Planta Med 2005; 71(2): 130-4.
  22. Tomoda M, Shimizu N, Kanari M, Gonda R, Arai S, Okuda Y. Characterization of two polysaccharides having activity on the reticuloendothelial system from the root of Glycyrrhiza uralensis. Chem Pharm Bull (Tokyo) 1990; 38(6): 1667-71.
  23. Hsiang CY, Lai IL, Chao DC, Ho TY. Differential regulation of activator protein 1 activity by glycyrrhizin. Life Sci 2002; 70(14): 1643-56.
Journal of Isfahan Medical School
Volume 32, Issue 298 - Serial Number 298
July and August 2014
Pages 1338-1346

Files

History

  • Receive Date: 16 March 2014
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

Share

How to cite

Statistics