Evaluation of anti-proliferative and cytotoxic effects of chlorogenic acid on breast cancer cell lines by real-time, label-free and high-throughput screening

Chlorogenic acid (CGA) is a major polyphenol in primary human diet, displaying a wide range of biological activities such as anti-oxidant, anti-inflammatory and anti-cancer effects. Several studies reported the chemopreventive effects of CGA on different types of cancers including breast cancer, which is the most common cancer among women worldwide. In this comprehensive study, we determined the anti-proliferative and cytotoxic effects of pure CGA on the phenotype of various breast cancer cell lines (MCF-7, SKBR-3, MDA-MB-231, MDA-MB-468 and BT- 20) with well-defined molecular classification and characteristics, in a time and dose dependent manner by using iCELLigence real-time and label-free cell analysis technology. Cells were plated on iCELLigence system-specific E-plate L8 and treated with CGA for 72 hours at concentrations ranging from 250 μM to 8 mM. Obtained data were analyzed by RTCA data analysis software 1.0 and IC50 values of 952±32.5 μM for MCF-7, 940±21.2 μM for SKBR-3, 590.5±10.6 μM for MDA-MB-231, 882.5±12.0 μM for MDA-MB-468 and 1095±121.6 μM for BT-20 cell lines were calculated at the end of 72-hour assay. Based on our findings, CGA did not have cytotoxic activity on breast cancer cell lines, and IC50 values and growth curves displayed similar anti-proliferative patterns with slight variation.

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[1] Cragg GM, Newman DJ. Natural products: A continuing source of novel drug leads. Biochim Biophys Acta - Gen Subj. 2013; 1830: 3670-3695.

[2] Shen B. A new golden age of natural products drug discovery. Cell. 2015; 163:1297-1300.

[3] Surh YJ. Cancer chemoprevention with dietary phytochemicals Nat Rev Cancer. 2003; 3: 768-780.

[4] Yin SY, Yang NS, Lin TJ. Phytochemicals approach for developing cancer immunotherapeutics. Front Pharmacol. 2017; 8: 386.

[5] Costa C, Tsatsakis A, Mamoulakis C, Teodoro M, Briguglio G, Caruso E, Tsoukalas D, Margina D, Dardiotis E, Kouretas D, Fenga C. Current evidence on the effect of dietary polyphenols intake on chronic diseases. Food Chem Toxicol. 2017; 110: 286-299.

[6] Kaur V, Kumar M, Kumar A, Kaur K, Dhillon VS, Kaur S. Pharmacotherapeutic potential of phytochemicals: Implications in cancer chemoprevention and future perspectives. Biomed Pharmacother. 2017; 97: 564-586.

[7] Hosseini AG, Ghorbani A. Cancer therapy with phytochemicals: evidence from clinical studies. Avicenna J Phytomed. 2015; 5: 84-97.

[8] Maru GB, Hudlikar RR, Kumar G, Gandhi K, Mahimkar MB. Understanding the molecular mechanisms of cancer prevention by dietary phytochemicals: From experimental models to clinical trials. World J Biol Chem 2016; 7: 88-99.

[9] Russo M, Spagnuolo C, Tedesco I, Russo GL. Phytochemicals in cancer prevention and therapy: Truth or dare? Toxins. 2010; 2 :517-551.

[10] Farah A, Monteiro M, Donangelo CM, Lafay S. Chlorogenic acids from green coffee extract are highly bioavailable in humans. J Nutr. 2008; 138: 2309–2315.

[11] Stalmach A, Crozier A, Clifford MN, Williamson G. Phytochemicals in coffee and the bioavailability of chlorogenic acids. In: Crozier A, Ashihara H, Tomás-Barbéran F. (Eds). Teas, cocoa and coffee: Plant secondary metabolites and health. Wiley-Blackwell, Oxford, UK, 2011, pp. 143–168.

[12] Naveed M, Hejazi V, Abbas M, Kamboh AA, Khan GJ, Shumzaid M, Ahmad F, Babazadeh D, FangFang X, Modarresi- Ghazani F, WenHua L, XiaoHui Z. Chlorogenic acid (CGA): A pharmacological review and call for further research. Biomed Pharmacother. 2017; 97: 67-74.

[13] Hunyadi A, Martins A, Hsieh T-J, Seres A, Zupkó I. Chlorogenic acid and rutin play a major role in the in vivo antidiabetic activity of Morus alba leaf extract on type II diabetic rats. PLoS One. 2012; 7 (11): e50619.

[14] dos Santos MD, Almeida MC, Lopes NP, de Souza GE. Evaluation of the anti-inflammatory, analgesic and antipyretic activities of the natural polyphenol chlorogenic acid. Biol Pharm Bull. 2006; 29: 2236–2240.

[15] Cho AS, Jeon SM, Kim MJ, Yeo J, Seo KI, Choi MS, Lee MK. Chlorogenic acid exhibits anti-obesity property and improves lipid metabolism in high-fat diet-induced-obese mice. Food Chem. Toxicol. 2010; 48: 937–943.

[16] Yamagata K, Izawa Y, Onodera D, Tagami M. Chlorogenic acid regulates apoptosis and stem cell marker-related gene expression in A549 human lung cancer cells. Mol Cell Biochem. 2017 [Epub ahead of print].

[17] Palmioli A, Ciaramelli C, Tisi R, Spinelli M, De Sanctis G, Sacco E, Airoldi C. Natural compounds in cancer prevention: Effects of coffee extracts and their main polyphenolic component, 5-O-Caffeoylquinic acid, on oncogenic Ras proteins. Chem Asian J. 2017; 12: 2457–2466.

[18] Xue N, Zhou Q, Ji M, Jin J, Lai F, Chen J, Zhang M, Jia J, Yang H, Zhang J, Li W, Jiang J, Chen X. Chlorogenic acid inhibits glioblastoma growth through repolarizating macrophage from M2 to M1 phenotype. Sci Rep. 2017; 7:39011.

[19] Lee J, Kim YS, Lee J, Heo SC, Lee KL, Choi SW, Kim Y. Walnut phenolic extract and its bioactive compounds suppress colon cancer cell growth by regulating colon cancer stemness. Nutrients. 2016; 8 (7) pii: E439.

[20] Shao P, Zhang JF, Chen XX, Sun PL. Microwave-assisted extraction and purification of chlorogenic acid from byproducts of Eucommia Ulmoides Oliver and its potential anti-tumor activity. J Food Sci Technol. 2015; 52: 4925-4934.

[21] Agar OT, Dikmen M, Ozturk N, Yilmaz MA, Temel H, Turkmenoglu FP. Comparative studies on phenolic composition, antioxidant, wound healing and cytotoxic activities of selected Achillea L. species growing in Turkey. Molecules. 2015; 20: 17976–18000.

[22] Méry B, Guy JB, Vallard A, Espenel S, Ardail D, Rodriguez-Lafrasse C, Rancoule C, Magné N. In vitro cell death determination for drug discovery: A landscape review of real issues. J Cell Death 2017; 10:1179670717691251.

[23] Kepp O, Galluzzi L, Lipinski M, Yuan J, Kroemer G. Cell death assays for drug discovery. Nat Rev Drug Discov. 2011; 10 :221-237.

[24] Ke N, Wang X, Xu X, Abassi YA. The xCELLigence system for real-time and label-free monitoring of cell viability. Methods Mol Biol. 2011; 740: 33–43.

[25] Kho D, MacDonald C, Johnson R, Unsworth CP, O’Carroll SJ, du Mez E, Angel CE, Graham ES. Application of xCELLigence RTCA biosensor technology for revealing the profile and window of drug responsiveness in real time. Biosensors. 2015; 5: 199–222.

[26] Singla AK, Garg A, Aggarwal D. Paclitaxel and its formulations. Int J Pharm. 2002; 235: 179–192.

[27] Mckeage K. Docetaxel: a review of its use for the first-line treatment of advanced castration-resistant prostate cancer. Drugs. 2012; 72: 1559–1577.

[28] Neve RM, Chin K, Fridlyand J, Yeh J, Baehner FL, Fevr T, Clark L, Bayani N, Coppe JP, Tong F, Speed T, Spellman PT, DeVries S, Lapuk A, Wang NJ, Kuo WL, Stilwell JL, Pinkel D, Albertson DG, Waldman FM, McCormick F, Dickson RB, Johnson MD, Lippman M, Ethier S, Gazdar A, Gray JW. A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. Cancer Cell. 2006; 10: 515–527.

[29] Shao W, Brown M. Advances in estrogen receptor biology: prospects for improvements in targeted breast cancer therapy. Breast Cancer Res. 2004; 6: 39-52.

[30] Holliday DL, Speirs V. Choosing the right cell line for breast cancer research. Breast Cancer Res. 2011; 13 (4): 215.

[31] Burdall SE, Hanby AM, Lansdown MRJ, Speirs V. Breast cancer cell lines: friend or foe? Breast Cancer Res. 2003; 5(2): 89-95.

[32] Mileo AM, Di Venere D, Linsalata V, Fraioli R, Miccadei S. Artichoke polyphenols induce apoptosis and decrease the invasive potential of the human breast cancer cell line MDA-MB231. J Cell Physiol. 2012; 227: 3301–3309..

[33] Deka S, Gorai S, Manna D, Trivedi V. Evidence of PKC binding and translocation to explain the anticancer mechanism of chlorogenic acid in breast cancer cells. Curr Mol Med. 2017; 17: 79–89.

[34] Suberu JO, Romero-Canelón I, Sullivan N, Lapkin AA, Barker GC. Comparative cytotoxicity of artemisinin and cisplatin and their interactions with chlorogenic acids in MCF7 breast cancer cells. ChemMedChem. 2014; 9: 2791– 2797.

[35] Naso LG, Valcarcel M, Roura-Ferrer M, Kortazar D, Salado C, Lezama L, Rojo T, González-Baró AC, Williams PA, Ferrer EG. Promising antioxidant and anticancer (human breast cancer) oxidovanadium(IV) complex of chlorogenic acid. Synthesis, characterization and spectroscopic examination on the transport mechanism with bovine serum albumin. J Inorg Biochem. 2014; 135: 86–99.

[36] Llorent-Martínez EJ, Zengin G, de Córdova MLF, Bender O, Atalay A, Ceylan R, Mollica A, Mocan A, Uysal S, Guler GO, Aktumsek A. Traditionally used Lathyrus species: Phytochemical composition, antioxidant activity, enzyme inhibitory properties, cytotoxic effects, and in silico studies of L. Czeczottianus and L. Nissolia. Front Pharmacol. 2017; 8:83.

[37] Picot MCN, Bender O, Atalay A, Zengin G, Loffredo L, Hadji-Minaglou F, Mahomoodally MF. Multiple pharmacological targets, cytotoxicity, and phytochemical profile of Aphloia theiformis (Vahl.) Benn. Biomed Pharmacother. 2017; 89: 342–350.