Biochemical profile, selective cytotoxicity and molecular effects of Calendula officinalis extracts on breast cancer cell lines
Keywords:apoptosis; breast cancer; Calendula officinalis; phenolic constituents; proliferation; selective cytotoxicity; volatile constituents
Calendula officinalis extracts have been known to possess anti-tumor properties, but questions regarding their mechanisms of action still need to be answered. Therefore, the present study aims to investigate the selective cytotoxicity, the biochemical profile and the corresponding molecular effects of two extracts of C. officinalis: flowers and leaves, against several breast cancer cell lines in vitro. Dry flowers and leaves were subjected to ultrasonication assisted extraction in methanol 70%. The phenolic and volatile profiles of the extracts, determined by HPLC-MS and nontargeted GC-MS, revealed high levels of specific phenolic acids, flavonols and coumarin and several volatile compounds, including mono- and sesquiterpenes, ketones, aldehydes and esters. Both extracts proved to possess selective cytotoxic activities against tumor cells in comparison to healthy endothelial cells, according to the MTT assay. The flower extract was superior in terms of both cytotoxicity and selectivity when compared to the leaf extract, in accordance to their biochemical profiles. The gene expression pattern for 10 genes of interest was evaluated by RT-qPCR. The expression level of several genes involved in apoptosis (BCL2, BAX, BBC3, ZMAT3), and cell cycle progression (NFkB, CCND1, STAT3) was modulated by the treatment with both extracts. Therefore, C. officinalis extracts proved to be rich in compounds characterized by cancer-related cytotoxicity and are capable of inducing selective cytotoxicity on breast cancer cell lines.
Ali F, Khan R, Khan AQ, Lateef MA, Maqbool T, Sultana S (2014). Assessment of augmented immune surveillance and tumor cell death by cytoplasmic stabilization of p53 as a chemopreventive strategy of 3 promising medicinal herbs in murine 2-stage skin carcinogenesis. Integrative Cancer Therapies 13(4):351-367.
Barajas-Farias LM, Perez-Carreon JI, Arce-Popoca E, Fattel-Fazenda S, Aleman-Lazarini L, Hernandez-Garcia S (2006). A dual and opposite effect of Calendula officinalis flower extract: chemoprotector and promoter in a rat hepatocarcinogenesis model. Planta Medica 72(3):217-221.
Bersani C, Xu LD, Vilborg A, Lui WO, Wiman KG (2014). Wig-1 regulates cell cycle arrest and cell death through the p53 targets FAS and 14-3-3sigma. Oncogene 33(35):4407-4417.
Bogdanova NS, Nikolaeva IS, Shcherbakova LI, Tolstova TI, Moskalenko N, Pershin GN (1970). Farmakologiia i toksikologiia [Study of antiviral properties of Calendula officinalis]. 33(3):349-355.
Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A (2018). Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians 68(6):394-424.
Butnariu M, Coradini CZ (2012). Evaluation of biologically active compounds from Calendula officinalis Flowers using spectrophotometry. Chemistry Central journal 6:35-43.
Chemat F, Rombaut N, Sicaire AG, Meullemiestre A, Fabiano-Tixier AS, Abert-Vian M (2017). Ultrasound assisted extraction of food and natural products. Mechanisms, techniques, combinations, protocols and applications. A review. Ultrasonics Sonochemistry 34:540-560.
Cruceriu D, Balacescu O, Rakosy E (2018). Calendula officinalis: potential roles in cancer treatment and palliative care. Integrative Cancer Therapies 17(4):1068-1078.
Dhawan D, Gupta J (2017). Comparison of different solvents for phytochemical extraction potential from Datura metel plant leaves. International Journal of Biological Chemistry 11(1):17-22.
Dhifi W, Bellili S, Jazi S, Bahloul N, Mnif W (2016). Essential Oils' chemical characterization and investigation of some biological activities: a critical review. Medicines 3(4):123-141.
dos Santos Junior HM, Oliveira DF, de Carvalho DA, Pinto JM, Campos VA, Mourao AR (2010). Evaluation of native and exotic Brazilian plants for anticancer activity. Journal of Natural Medicines 64(2):231-238.
Efstratiou E, Hussain AI, Nigam PS, Moore JE, Ayub MA, Rao JR (2012). Antimicrobial activity of Calendula officinalis petal extracts against fungi, as well as Gram-negative and Gram-positive clinical pathogens. Complementary Therapies in Clinical Practice 18(3):173-176.
Fantini M, Benvenuto M, Masuelli L, Frajese GV, Tresoldi I, Modesti A, Bei R (2015). In vitro and in vivo antitumoral effects of combinations of polyphenols, or polyphenols and anticancer drugs: perspectives on cancer treatment. International Journal of Molecular Sciences 16(5):9236-9282.
Frankic T, Salobir K, Salobir J (2009). The comparison of in vivo antigenotoxic and antioxidative capacity of two propylene glycol extracts of Calendula officinalis (marigold) and vitamin E in young growing pigs. Journal of Animal Physiology and Animal Nutrition 93(6):688-694.
Gazim ZC, Rezende CM, Fraga SR, Svidzinski TI, Cortez DA (2008). Antifungal activity of the essential oil from Calendula officinalis L. (Asteraceae) growing in Brazil. Brazilian Journal of Microbiology [Publication of the Brazilian Society for Microbiology] 39(1):61-63.
Greay SJ, Hammer KA (2015). Recent developments in the bioactivity of mono- and diterpenes: anticancer and antimicrobial activity. Phytochemistry Reviews 14(1):6.
Holliday DL, Speirs V (2011). Choosing the right cell line for breast cancer research. Breast Cancer Research BCR 13(4):215-227.
Horneber M, Bueschel G, Dennert G, Less D, Ritter E, Zwahlen M (2012). How many cancer patients use complementary and alternative medicine: a systematic review and metaanalysis. Integrative Cancer Therapies 11(3):187-203.
Hu Z, Yang X, Ho PC, Chan SY, Heng PW, Chan E (2005). Herb-drug interactions: a literature review. Drugs 65(9):1239-1282.
Hui LM, Zhao GD, Zhao JJ (2015). Delta-cadinene inhibits the growth of ovarian cancer cells via caspase-dependent apoptosis and cell cycle arrest. International Journal of Clinical and Experimental Pathology 8(6):6046-6056.
Iqbal J, Abbasi B, Mahmood T, Kanwal S, Ali B, Shah SA, Khalil AT (2017). Plant-derived anticancer agents: A green anticancer approach. Asian Pacific Journal of Tropical Biomedicine 7(12):21.
Jimenez-Medina E, Garcia-Lora A, Paco L, Algarra I, Collado A, Garrido F (2006). A new extract of the plant Calendula officinalis produces a dual in vitro effect: cytotoxic anti-tumor activity and lymphocyte activation. BMC Cancer 6:119-132.
Kashyap D, Mittal S, Sak K, Singhal P, Tuli HS (2016). Molecular mechanisms of action of quercetin in cancer: recent advances. Tumour Biology 37(10):12927-12939.
Kaškonienė V, Kaškonas P, Jalinskaitė M, Maruška A (2011). Chemical Composition and chemometric analysis of variation in essential oils of Calendula officinalis L. during vegetation stages. Chromatographia 73(1):163-169.
Lewandowska H, Kalinowska M, Lewandowski W, Stepkowski TM, Brzoska K (2016). The role of natural polyphenols in cell signaling and cytoprotection against cancer development. The Journal of Nutritional Biochemistry 32:1-19.
Li L, Zhao P, Hu J, Liu J, Liu Y, Wang Z (2015). Synthesis, in vitro and in vivo antitumor activity of scopoletin-cinnamic acid hybrids. European Journal of Medicinal Chemistry 93:300-307.
Livak KJ, Schmittgen TD (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25(4):402-408.
Matic IZ, Juranic Z, Savikin K, Zdunic G, Nadvinski N, Godevac D (2013). Chamomile and marigold tea: chemical characterization and evaluation of anticancer activity. Phytotherapy Research 27(6):852-858.
Matysik G, Wojciak-Kosior M, Paduch R (2005). The influence of Calendulae officinalis flos extracts on cell cultures, and the chromatographic analysis of extracts. Journal of pharmaceutical and biomedical analysis 38(2):285-292.
Mehta D, Rastogi P, Kumar AK (2012). Review on Pharmacological Update: Calendula officinalis Linn. Inventi Impact: Planta Activa 2012(4):195-204.
Miguel M, Barros L, Pereira C, Calhelha RC, Garcia PA, Castro M (2016). Chemical characterization and bioactive properties of two aromatic plants: Calendula officinalis L. (flowers) and Mentha cervina L. (leaves). Food and Function 7(5):2223-2232.
Mouhid L, Gomez de Cedron M, Vargas T, Garcia-Carrascosa E, Herranz N, Garcia-Risco M (2018). Identification of antitumoral agents against human pancreatic cancer cells from Asteraceae and Lamiaceae plant extracts. BMC Complementary and Alternative Medicine 18(1):254-262.
Nakamura ES, Kurosaki F, Arisawa M, Mukainaka T, Okuda M, Tokuda H (2002). Cancer chemopreventive effects of constituents of Caesalpinia ferrea and related compounds. Cancer Letters 177(2):119-124.
Newman DJ, Cragg GM (2016). Natural products as sources of new drugs from 1981 to 2014. Journal of Natural Products 79(3):629-661.
Nichenametla SN, Taruscio TG, Barney DL, Exon JH (2006). A review of the effects and mechanisms of polyphenolics in cancer. Critical Reviews in Food Science and Nutrition 46(2):161-183.
Okoh OO, Sadimenko AA, Afolayan AJ (2007). The effects of age on the yield and composition of the essential oils of Calendula officinalis. Journal of Applied Sciences 7(23):3806-3811.
Olaku O, White JD (2011). Herbal therapy use by cancer patients: a literature review on case reports. European Journal of Cancer 47(4):508-514.
Olennikov DN, Kashchenko NI, Chirikova NK, Akobirshoeva A, Zilfikarov IN, Vennos C (2017). Isorhamnetin and quercetin derivatives as anti-acetylcholinesterase principles of marigold (Calendula officinalis) flowers and preparations. International Journal of Molecular Sciences 18(8):225-264.
Parente LM, Lino Junior Rde S, Tresvenzol LM, Vinaud MC, de Paula JR, Paulo NM (2012). Wound healing and anti-inflammatory effect in animal models of Calendula officinalis L. growing in Brazil. Evidence-Based Complementary and Alternative Medicine eCAM 2012:375671.
Peto R, Davies C, Godwin J, Gray R, Pan HC, Clarke M (2012). Comparisons between different polychemotherapy regimens for early breast cancer: meta-analyses of long-term outcome among 100,000 women in 123 randomised trials. Lancet 379(9814):432-444.
Petrović L, Lepojević Z, Sovilj V, Adamović DVT (2010). Composition of essential oil obtained from tubular, head and ligulate flowers of Calendula officinalis L. by steam distillation of plant material and CO2 extracts. Journal of Essential Oil Research 22(2):4.
Preethi KC, Siveen KS, Kuttan R, Kuttan G (2010). Inhibition of metastasis of B16F-10 melanoma cells in C57BL/6 mice by an extract of Calendula officinalis L flowers. Asian Pacific Journal of Cancer Prevention 11(6):1773-1779.
Rigane G, Younes SB, Ghazghazi H, Salem R (2013). Investigation into the biological activities and chemical composition of Calendula officinalis L. growing in Tunisia. International Food Research Journal 20(6):3001-3008.
Saghatchian M, Bihan C, Chenailler C, Mazouni C, Dauchy S, Delaloge S (2014). Exploring frontiers: use of complementary and alternative medicine among patients with early-stage breast cancer. Breast 23(3):279-285.
Sulaiman SF, Sajak AAB, Ooi KL, M. SE (2011). Effect of solvents in extracting polyphenols and antioxidants of selected raw vegetables. Journal of Food Composition and Analysis 24(4-5):506-516.
Turkez H, Togar B, Tatar A, Geyıkoglu F, Hacımuftuoglu A (2014). Cytotoxic and cytogenetic effects of α-copaene on rat neuron and N2a neuroblastoma cell lines. Biologia 69(7):936-943.
Varlijen J (1989). Structural analysis of rhamnoarabinogalactans and arabinogalactans with immunostimulating activity from Calendula officinalis. Phytochemistry 28:2379-2384.
Wegiera M, Smolarz HD, Jedruch M, Korczak M, Kopron K (2012). Cytotoxic effect of some medicinal plants from Asteraceae family on J-45.01 leukemic cell line--pilot study. Acta Poloniae Pharmaceutica 69(2):263-268.
Wu Q, Kroon PA, Shao H, Needs PW, Yang X (2018). Differential effects of quercetin and two of its derivatives, isorhamnetin and isorhamnetin-3-glucuronide, in inhibiting the proliferation of human breast-cancer MCF-7 cells. Journal of Agricultural and Food Chemistry 66(27):7181-7189.
Yamagata K, Izawa Y, Onodera D, Tagami M (2018). Chlorogenic acid regulates apoptosis and stem cell marker-related gene expression in A549 human lung cancer cells. Molecular and Cellular Biochemistry 441(1-2):9-19.
How to Cite
Open Access Journal:
The journal allows the author(s) to retain publishing rights without restriction. Users are allowed to read, download, copy, distribute, print, search, or link to the full texts of the articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author.