A study on the targeting of ceramide metabolism by (-)-epicatechin gallate, catechin and quercetin in A-549 lung cancer cell line

Mashhadi Akbar Boojar, Masoud

doi:10.29252/nbr.6.3.275

A study on the targeting of ceramide metabolism by (-)-epicatechin gallate, catechin and quercetin in A-549 lung cancer cell line

Author

Masoud Mashhadi Akbar Boojar

https://doi.org/10.29252/nbr.6.3.275

Abstract

Catechin, epicatechin gallate (ECG) and quercetin, as bioactive flavonoids, have been shown to possess anticarcinogenic effects. Ceramide plays an important role in killing tumor cells. Accordingly, the aim of this study was to clarify the involvement of these compounds in ceramide metabolism in A549 cancerous cell line. Spectrophotometer, cell culture and HPLC methods were used. Cell viability index showed different potential of cytotoxicity effect for each of the studied agents, among which ECG was more potent. This index decreased significantly over 100 to 250 µM concentrations of treatment with respect to control. Cell treatments also caused considerable increase in ceramide level within cells in a dose-dependent manner. Sphingomyelinase activity increased significantly in treatment with quercetin and catechin. There was significant inhibition in acid ceramidase activity of cell extract in response to each of the three compounds, particularly over 100 µM in comparison with control. Data also showed no significant variation in glycosyl ceramide synthase activity in treated cells with quercetin, whereas the activity decreased significantly by Catechin and/or ECG. It is our conviction that different effects on ceramide metabolism enzymes may be related to various chemical groups on the common structure of the studied compounds. Due to structure-function relationship, these compounds had different effects on ceramide generation. Elevation in ceramide content in A549 cancer cell line induced apoptosis, which led to anti-cancerous effects, as observed in this study.

Keywords

A549 cell line

catechin

ceramide

epicatechin gallate

quercetin

20.1001.1.24236330.1398.6.3.2.1

Aiping, B., Cungui, M., Russell, W.J., Zdzislaw, M.S., Alicja, B. and Yusuf, A.H. 2017. Anticancer actions of lysosomally targeted inhibitor, LCL521, of acid ceramidase. – PLoS ONE 12: 1-10.
Anna, K., Anna, R., Michal, G., Jerzy, S. and Grazyna, S. 2014. Structure and antioxidant activity of polyphenols derived from Propolis. – Molecules 19: 78-101.
Benavente-Garcia, O. and Castillo, J. 2008. Update on uses and properties of citrus flavonoids: new findings in anticancer, cardiovascular, and anti-inflammatory activity. – J. Agric. Food. Chem. 56: 6185-6205.
Carlos, M.P., Carol, W., Arran, K. T., Peter, M., Graeme, C. and James, M. 2016. Antitumour activity of the novel flavonoid Oncamex in preclinical breast cancer models. – British J. Cancer 114: 905-916.
Chang, H., Mi, M., Ling, W., Zhu, J., Zhang, Q., Wei, N.A., Zhou, Y., Tang, Y., Yu, X. and Zhang, T.I.N. 2010. Structurally related anticancer activity of flavonoids: involvement of reactive oxygen species generation. – J. Food Biochem. 34:1-14.
Chou, C.C., Yang, J.S, Lu, H.F., Ip, S.W., Lo, C., Wu, C.C., Lin, J.P., Tang, N.Y., Chung, J.G. and Chou, M.J. 2010. Quercetin-mediated cell cycle arrest and apoptosis involving activation of a caspase cascade through the mitochondrial pathway in human breast cancer MCF-7 cells. – Arch. Pharm. Res. 33: 1181-1191.
Elisa, R.C. Paolo, N. Francisco, M. and Iván L.M. 2017. The enzymatic sphingomyelin to ceramide conversion increases the shear membrane viscosity at the air-water interface. – Adv. Colloid Interface Sci. 247: 555-560.
Galati, G.O'. Brien, P.J. 2004. Potential toxicity of flavonoids and other dietary phenolics: significance for their chemopreventive and anticancer properties. – Free Radic. Biol. Med. 37: 287-303.
Geraldine, R., Christophe, S., Benoit, L., Hervé, S., Hamid, M., Hassan, E.L., Btaouri, S. and Dedieu, L.M. 2009. De novo ceramide synthesis is responsible for the anti-tumor properties of camptothecin and doxorubicin in follicular thyroid carcinoma. – Int. J. Biochem. Cell Biol. 41: 1165-1172.
Guohua, H., Lei, Z., Yefei, R., Xiaoling, N. and Yihong Sun. 2014. Downstream carcinogenesis signaling pathways by green tea polyphenols: A translational perspective of chemoprevention and treatment for cancers. – Curr. Drug Metabol. 15: 14-22.
Han, D.H., Jeong, J.I.N.H. and Kim, J.H.E.E. 2009. Anti-proliferative and apoptosis induction activity of green tea polyphenols on human promyelocytic leukemia HL-60 cells. – Anticancer Res. 29: 1417-1421.
Jeong, J.H., An, J.Y., Kwon, Y.T., Rhee, J.G. and Lee, Y.J. 2009. Effects of low dose quercetin: cancer cell-specific inhibition of cell cycle progression. – J. Cell Biochem. 106: 73-82.
Ki Duk, P., Sul, Gi, L., Sung, U.K.K., Sung Han, K., Won Suck, S., Sung Jin, C. and Do Hyeon, J. 2004. Anticancer activity of 3-O-acyl and alkyl-(-)-epicatechin derivatives. – Bioorg. Med. Chem. 14: 5189-5192.
Kim, M.E., Ha, T.K., Yoon, J.H. and Lee, J.S. 2014. Myricetin induces cell death of human colon cancer cells via BAX/BCL2-dependent pathway. – Anticancer Res. 34: 701-6.
Krishna, P.B., Burkhard, K., Andrea, H. and Christoph, A. 2013. Effective inhibition of acid and neutral ceramidases by novel B-13 and LCL-464 analogues. – Bioorg. Med. Chem. 21: 874-882.
Kumar, G. and Baojun, X. 2018. Telomerase inhibitors from natural products and their anticancer potential. ‒ Int. J. Mol. Sci. 19: 2-26.
Lafont, E., Dupont, R., Andrieu-Abadie, N., Okazaki, T. and Schulze-Osthoff, K. 2012. Ordering of ceramide formation and caspase-9 activation in CD95L-induced Jurkat leukemia T cell apoptosis. – Biochim. Biophys. Acta. 1821:684-693.
Leah, J. Siskind. 2005. Mitochondrial Ceramide and the Induction of Apoptosis. – J. Bioenerg. Biomembr. 37: 143-153.
Lee, Y.K, Park, S.Y, Kim, Y.M., Lee, W.S. and Park, O.J. 2009. AMP kinase/cyclooxygenase-2 pathway regulates proliferation and apoptosis of cancer cells treated with quercetin. ‒ Exp. Mol. Med. 41: 201-207.
Lin, C.F, Chen, C.L. andLin, Y.S. 2006. Ceramide in apoptotic signaling and anticancer therapy. ‒ Curr. Med. Chem. 13:1609-1616.
Lotito, S.B. and Frei, B. 2006. Consumption of Flavonoid-rich foods and increased plasma antioxidant capacity in human: cause, consequence or epiphenomenon. – Free Radic. Biol. Med. 41: 1727-1746.
Maggioni, D., Nicolini, G., Rigolio, R., Biffi, L., Pignataro, L., Gaini, R. and Garavello, W. 2014. Myricetin and naringenin inhibit human squamous cell carcinoma proliferation and migration in vitro. ‒ Nutr. Cancer 66: 1257-67.
Maurya P.K. and Rizvi, S.I. 2009. Protective role of tea catechins on erythrocytes subjected to oxidative stress during human aging. – Nat. Prod. Res. 23: 1072-1079.
Mendosa-Wilson, A.M., and Glossman-Mitnik, D. 2006. Theoretical study of the molecular properties and chemical reactivity of (+) - catechin and (-)-epicatechin related to their antioxidant ability. – J. Mol. Struct. 761: 97-106.
Mi Sun, K., Kyong Hoon, A., Seok Kyun, K., Hyung, J.J., Jung Eun, J., Jong Min, C., Kwang Mook, J., Sung Yun, J. and Dae Kyong, K. 2010. Hypoxia-induced neuronal apoptosis is mediated by de novo synthesis of ceramide through activation of serine palmitoyltransferase. ‒ Cell. Signal. 22: 610-618.
Moretti, E. Mazzi, L., Terzuoli, G., Bonechi, C., Lacoponi, F., Martini, S., Rossi, C. and Collodel, G. 2012. Effect of quercetin, rutin, naringenin and epicatechin on lipid peroxidation on lipid peroxidation induced in human sperm. – Reprod. Toxicol. 34: 651-657.
Munawar, A., Farhan, S., Faqir, M., Anjum, M.A., Tabussam, T. and Muhammad Shakeel, B. 2017. Natural polyphenols: An overview. ‒ Int. J. Food Prop. 20:1689-1699.
Negrão, R., Costa, R., Duarte, D., Taveira, G.T., Mendanha, M. and Moura, L. 2010. Angiogenesis and inflammation signaling are targets of beer polyphenols on vascular cells. – J. Cell Biochem. 111: 1270-1279.
Ogretmen, B. and Hannun, Y.A. 2004. Biologically active sphingolipids in cancer pathogenesis and treatment. – Nat. Rev. Cancer 4: 604-616.
Sak, K. 2014. Dependence of DPPH radical scavenging activity of dietary flavonoid quercetin on reaction environment. – Mini Rev. Med. Chem. 14: 494-504.
Shyi-Neng, L., Ya-Siou, H. and Chi-Tang, H. 2014. Flavonoid compositions and antioxidant activity of calamondin extracts prepared using different solvents. – J. Food Drug Anal. 22: 290-295.
Smarajit, M., Aarifa, N., Nandita, M., Ritesh, P. and Tamal Kanti, G. 2019. Flavonoids green tea against oxidant stress and inflammation with related human diseases. – Clin. Nutr. Experi. 24: 1-14.
Soares, R., Balogh, G., Guo, S., Gartner, F., Russo, J. and Schmitt, F. 2004. Evidence for the notch signaling pathway on the role of estrogen in angiogenesis. – Mol. Endocrinolgy 18: 2333-2343.
Subhadra, R. 2016. Cancer, a preventable disease of the modern age-an overview from the Indian perspective. – Intl. J. Bioinf. Biol. Sci. 4: 1-4.
Teillet, F., Boumendjel, A., Boutonnat, J. and Ronot, X. 2008. Flavonoids as RTK inhibitors and potential anticancer agents. – Med. Res. Rev. 28: 715-745.
Wang, H., Maurer, B.J., Reynolds, C.P. and Cabot, M.C. 2001. N-(4-hydroxyphenyl) retinamide elevates ceramide in neuroblastoma cell lines by coordinate activation of serine palmitoyltransferase and ceramide synthase. – Cancer Res. 61: 5102-5105.
Williams, R.J., Spencer, J.P. and Rice-Evans, C. 2004. Flavonoids: antioxidants or signalling molecules? – Free Radic. Biol. Med. 36: 838-849.
Xingxuan, H., Arie, D., Shimon, G. and Edward, H.S. 2005. Simultaneous quantitative analysis of ceramide and sphingosine in mouse blood by naphthalene-2,3-dicarboxyaldehyde derivatization after hydrolysis with ceramidase. – Anal. Biochem. 340: 113-122.
Yasuhiro, H., Yasuhiro, H., Keishi, S., Nozomu, O. and Makoto, I. 2005. A sensitive and reproducible assay to measure the activity of glucosylceramide synthase and lactosylceramide synthase using HPLC and Fluorescence substrates. – Anal. Biochem. 345: 181-186.
Zhou, M., Diwu, Z., Panchuk-Voloshina, N. and Haugland, R.P. 1997. A stable nonfluorescent derivative of resorufin for the fluorometric determination of trace hydrogen peroxide: applications in detecting the activity of phagocyte NADPH oxidase and other oxidases. – Anal. Biochem. 253: 62-68.