Abstract
Breast cancer is the most diagnosed invasive cancer and the second leading cause of cancer in women. Standard medicinal protocol for treatment of this disease implies surgery and use of chemotherapeutic drugs. It is known that these drugs are associated with resistance, cancer relapse, and numerous side effects. Researchers give great importance to natural bioactive compounds because of their conformed health beneficial effects. Ocimum minimum L. is a well-known plant in folk medicine for treating various diseases and conditions, whit a large specter of pharmaceutical activities. The aim of this study is to evaluate anticancer properties and impact on fatty acid synthesis process under the influence of O. minimum L. flowers in triple-negative breast cancer cell lines (MDA-MB-231 and MDA-MB-468) and normal fetal fibroblasts (MRC-5). The impact of O. minimum L. on fatty acid synthesis process in breast cancer cells has been evaluated through cytotoxic activity by MTT assay, pro-apoptotic activity by AO/EB staining, concentrations of redox status parameters (O2.-, lipid peroxidation, GSH) by colorimetric methods. The expression of genes involved in fatty acid synthesis (ACLY, FASN, MAGL, HIF-1α) were determined by the qPCR method, while the protein expression (FASN) was analyzed by immunocytochemistry. Based on the obtained results, synergic action between bioactive compounds identified from O. minimum L. showed cytotoxic, proapoptotic, and prooxidant effects on breast cancer cell lines, as well as suppression of genes and protein (FASN) involved in fatty acid synthesis process. This plant showed significant pharmacological effects whit potential use as nutritional supplement in anticancer therapy.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- Ocimum minimum L.:
-
O. minimum L.
- FAs:
-
Fatty acid synthesis
- ACLY:
-
ATP-citrate lyase
- Acetyl-CoA:
-
acetyl-coenzyme A
- ACC:
-
Acetyl-CoA carboxylase
- FASN:
-
Fatty acid synthase
- MAGL:
-
Monoacylglycerol lipase
- DMEM:
-
Dulbecco’s Modified Eagle Medium
- HPLC:
-
High-performance liquid chromatography
- MTT:
-
3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
- IC50:
-
The half maximal inhibitory concentration
- AO/EB:
-
Acridine orange/ethidium bromide
- NBT:
-
Nitroblue tetrazolium
- MDA:
-
Malondialdehyde
- TBARS:
-
Thiobarbituric acid reactive substances
- GSH:
-
Reduced glutathione
- ROS:
-
Reactive oxygen species
- HIF1α:
-
hypoxia-inducible factor-1α
References
Auclair C, Voisin E, 1985. Nitroblue tetrazolium reduction. In: Greenwald RA (ed) Handbook of methods for oxygen radical research. Boka Raton: CRC Press, p 123–132
Baker MA, Cerniglia GJ, Zaman A (1990) Microtiter plate assay for the measurement of glutathione and glutathione disulfide in large numbers of biological samples. Anal Biochem 190(2):360–365. https://doi.org/10.1016/0003-2697(90)90208-q
Baskić D, Popović S, Ristić P, Arsenijević NN (2006) Analysis of cycloheximide-induced apoptosis in human leukocytes: fluorescence microscopy using annexin V/propidium iodide versus acridin orange/ethidium bromide. Cell Biol Int 30(11):924–932. https://doi.org/10.1016/j.cellbi.2006.06.016
Brusselmans K, Vrolix R, Verhoeven G, Swinnen JV (2005) Induction of cancer cell apoptosis by flavonoids is associated with their ability to inhibit fatty acid synthase activity. J Biol Chem 280(7):5636–5645. https://doi.org/10.1074/jbc.M408177200
Buege JA, Aust SD (1987) Microsomal lipid peroxidation. Methods Enzymol 52:302–310. https://doi.org/10.1016/s0076-6879(78)52032-6
Chen M, Huang J (2019) The expanded role of fatty acid metabolism in cancer: new aspects and targets. Precision Clin Med 2(3):183–191. https://doi.org/10.1093/pcmedi/pbz017
Chomczynski P, Sacchi N (1987) Single-step method of RNА isolation by acid guanidinium thiocyanatephenol-chloroform extraction. Anal Biochem 162:156–159. https://doi.org/10.1006/abio.1987.9999
Currie E, Schulze A, Zechner R, Walther TC, Farese RV Jr (2013) Cellular fatty acid metabolism and cancer. Cell Metab 18(2):153–161. https://doi.org/10.1016/j.cmet.2013.05.017
DeBerardinis RJ (2008) Is cancer a disease of abnormal cellular metabolism? New angles on an old idea. Genet Med 10(11):767–777. https://doi.org/10.1097/GIM.0b013e31818b0d9b
Flegkas A, Milosević Ifantis T, Barda C, Samara P, Tsitsilonis O, Skaltsa H (2019) Antiproliferative activity of (−)-Rabdosiin isolated from Ocimum sanctum L. Medicines (Basel) 6(1):37. https://doi.org/10.3390/medicines6010037
Forman HJ, Davies KJ, Ursini F (2014) How do nutritional antioxidants really work: nucleophilic tone and Para-hormesis versus free radical scavenging in vivo. Free Radic Biol Med 66:24–35. https://doi.org/10.1016/j.freeradbiomed.2013.05.045
Grayer RJ, Vieira RF, Price AM, Kite GC, Simon JE, Paton AJ (2004) Characterization of cultivars within species of Ocimum by exudate flavonoid profiles. Biochem Syst Ecol 32(10):901–913. https://doi.org/10.1016/j.bse.2004.04.002
Grbović F, Stanković MS, Ćurčić M, Đorđević N, Šeklić D, Topuzović M, Marković S (2013) In vitro cytotoxic activity of Origanum vulgare L. on HCT-116 and MDA-MB-231 cell lines. Plants 2(3):371–378. https://doi.org/10.3390/plants2030371
Haffty BG, Yang Q, Reiss M, Kearney T, Higgins SA, Weidhaas J, Harris L, Hait W, Toppmeyer D (2006) Locoregional relapse and distant metastasis in conservatively managed triple negative early-stage breast cancer. J Clin Oncol 24(36):5652–5657. https://doi.org/10.1200/JCO.2006.06.5664
Higuchi H, Bronk SF, Takikawa Y, Werneburg N, Takimoto R, El-Deiry W, Gores GJ (2001) The bile acid glycochenodeoxycholate induces TRAIL-receptor 2/ DR5 expression and apoptosis. J Biol Chem 276(42):38610–38618. https://doi.org/10.1074/jbc.M105300200
Huang WY, Cai YZ, Zhang Y (2010) Natural phenolic compounds from medicinal herbs and dietary plants: potential use for cancer prevention. Nutr Cancer 62(1):1–20. https://doi.org/10.1080/01635580903191585
Iriti M, Kubina R, Cochis A, Sorrentino R, Varoni EM, Kabała-Dzik A, Azzimonti B, Dziedzic A, Rimondini L, Wojtyczka RD (2017) Rutin, a quercetin glycoside, restores Chemosensitivity in human breast Cancer cells. Phytother Res 31(10):1529–1538. https://doi.org/10.1002/ptr.5878
Jakovljević D, Topuzović M, Stanković M (2019) Nutrient limitation as a tool for the induction of secondary metabolites with antioxidant activity in basil cultivars. Ind Crop Prod 138:111462. https://doi.org/10.1016/j.indcrop.2019.06.025
Jamshidi-Kia F, Lorigooini Z, Amini-Khoei H (2018) Medicinal plants: past history and future perspective. J Herbmed Pharmacol 7(1):1–7. https://doi.org/10.15171/jhp.2018.01
Kumar S, Pandey AK (2013) Chemistry and biological activities of flavonoids: an overview. Sci World J 162750. https://doi.org/10.1155/2013/162750
Kyselova Z (2011) Toxicological aspects of the use of phenolic compounds in disease prevention. Interdiscip Toxicol 4(4):173–183. https://doi.org/10.2478/v10102-011-0027-5
Lee DH, Lee YJ (2008) Quercetin suppresses hypoxia-induced accumulation of hypoxia-inducible factor-1alpha (HIF-1alpha) through inhibiting protein synthesis. J Cell Biochem 105(2):546–553. https://doi.org/10.1002/jcb.21851
Leung HW, Lin CJ, Hour MJ, Yang WH, Wang MY, Lee HZ (2007) Kaempferol induces apoptosis in human lung non-small carcinoma cells accompanied by an induction of antioxidant enzymes. Food Chem Toxicol 45(10):2005–2013. https://doi.org/10.1016/j.fct.2007.04.023
Li SZ, Li K, Zhang JH, Dong Z (2013) The effect of quercetin on doxorubicin cytotoxicity in human breast cancer cells. Anti Cancer Agents Med Chem 13(2):352–355. https://doi.org/10.2174/1871520611313020020
Listenberger LL, Han X, Lewis SE, Cases S, Farese RV, Ory DS, Schaffer JE (2003) Triglyceride accumulation protects against fatty acid-induced lipotoxicity. Proc Natl Acad Sci U S A 100(6):3077–3082. https://doi.org/10.1073/pnas.0630588100
Li-Weber M (2009) New therapeutic aspects of flavones: the anticancer properties of Scutellaria and its main active constituents Wogonin, Baicalein and Baicalin. Cancer Treat Rev 35:57–68. https://doi.org/10.1016/j.ctrv.2008.09.005
Lushchak VI (2012) Glutathione homeostasis and functions: potential targets for medical interventions. J Amino Acids 2012:736837. https://doi.org/10.1155/2012/736837
Mahvi DA, Liu R, Grinstaff MW, Colson YL, Raut CP (2018) Local Cancer recurrence: the realities, challenges, and opportunities for new therapies. CA Cancer J Clin 68(6):488–505. https://doi.org/10.3322/caac.21498
Makkar HPS (2000) Quantification of tannins in tree foliage: a laboratory manual. Vienna: FAO/IAEA Edition
Matés JM, Segura JA, Alonso FJ, Márquez J (2008) Intracellular redox status and oxidative stress: implications for cell proliferation, apoptosis, and carcinogenesis. Arch Toxicol 82(5):273–299. https://doi.org/10.1007/s00204-008-0304-z
Menendez JA, Lupu R (2004) Fatty acid synthase-catalyzed de novo fatty acid biosynthesis: from anabolic-energy-storage pathway in normal tissues to jack-of-all-trades in cancer cells. Archivum Immunol Ther Exp 52(6):414–426
Milutinović M, Stanković M, Cvetković D, Maksimović V, Šmit B, Pavlović R, Marković S (2015) The molecular mechanisms of apoptosis induced by Allium flavum L. and synergistic effects with new-synthesized Pd(II) complex on colon cancer cells. J Food Biochem 39:238–250. https://doi.org/10.1111/jfbc.12123
Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65(1–2):55–63. https://doi.org/10.1016/0022-1759(83)90303-4
Nikodijević DD, Jovankić JV, Cvetković DM, Anđelković MZ, Nikezić AG, Milutinović MG (2021) L-amino acid oxidase from snake venom: biotransformation and induction of apoptosis in human colon cancer cells. Eur J Pharmacol 910:174466. https://doi.org/10.1016/j.ejphar.2021.174466
Quettier D, Greissier B, Vasseur J, Dine T, Brunet C, Luyckx M, Cazin M, Cazin JC, Bailleul F, Trotin F (2000) Phenolic compounds and antioxidant activities of buckwheat (Fagopyrum esculentum Moench) hulls and flour. J Ethnopharmacol 72(1–2):35–42. https://doi.org/10.1016/S0378-8741(00)00196-3
Sak K (2014) Cytotoxicity of dietary flavonoids on different human cancer types. Pharmacogn Rev 8(16):122–146. https://doi.org/10.4103/0973-7847.134247
Schmittgen T, Livak K (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 3:1101–1108. https://doi.org/10.1038/nprot.2008.73
Tian WX, Ma XF, Zhang SY, Sun YH, Li BH (2011) Fatty acid synthase inhibitors from plants and their potential application in the prevention of metabolic syndrome. Clin Oncol Cancer Res 8:1–9. https://doi.org/10.1007/s11805-011-0550-3
Torres RG, Casanova L, Carvalho J, Marcondes MC, Costa SS, Sola-Penna M, Zancan P (2018) Ocimum basilicum but not Ocimum gratissimum present cytotoxic effects on human breast cancer cell line MCF-7, inducing apoptosis and triggering mTOR/Akt/p70S6K pathway. J Bioenerg Biomembr 50:93–105. https://doi.org/10.1007/s10863-018-9750-3
Valli A, Rodriguez M, Moutsianas L, Fischer R, Fedele V et al (2015) Hypoxia induces a lipogenic cancer cell phenotype via HIF1α-dependent and -independent pathways. Oncotarget 6:1920–1941. https://doi.org/10.18632/oncotarget.3058
Vieira PR, de Morais SM, Bezerra FH, Ferreira PAT, Oliveira ÍR, Silva MGDV (2014) Chemical composition and antifungal activity of essential oils from Ocimum species. Ind Crop Prod 55:267–271. https://doi.org/10.1016/j.indcrop.2014.02.032
Wagner H, Ulrich-Merzenich G (2009) Synergy research: approaching a new generation of phytopharmaceuticals. Phytomedicine 16(2–3):97–110. https://doi.org/10.1016/j.phymed.2008.12.018
Wang H, Khor TO, Shu L, Su ZY, Fuentes F, Lee JH, Kong AT (2012) Plants vs. cancer: a review on natural phytochemicals in preventing and treating cancers and their druggability. Anti Cancer Agents Med Chem 12(10):1281–1305. https://doi.org/10.2174/187152012803833026
Wootton-Beard P, Ryan L (2011) A beetroot juice shot is a significant and convenient source of bioaccessible antioxidants. J Funct Foods 3(4):329–334. https://doi.org/10.1016/j.jff.2011.05.007
** D, Yijiao P, **gduo Z, **aoyong L, **ng Z, Zhizhong X, Guotao T (2019) Anticancer activity of natural flavonoids: inhibition of HIF-1α signaling pathway. Curr Org Chem 23(26):2945–2959. https://doi.org/10.2174/1385272823666191203122030
Zhou D, Shao L, Spitz DR (2014) Reactive oxygen species in normal and tumor stem cells. Adv Cancer Res 122:1–67. https://doi.org/10.1016/B978-0-12-420117-0.00001-3
Žižić JB, Vuković NL, Jadranin MB, Anđelković BD, Tešević VV, Kacaniova MM, Sukdolak SB, Marković SD (2013) Chemical composition, cytotoxic and antioxidative sctivities of etanolic extracts of propolis on HCT-116 cell line. J Sci Food Agric 93(12):3001–3009. https://doi.org/10.1002/jsfa.6132
Zorov DB, Juhaszova M, Sollott SJ (2014) Mitochondrial reactive oxygen species (ROS) and ROS-induced ROS release. Physiol Rev 94(3):909–950. https://doi.org/10.1152/physrev.00026.2013
Acknowledgments
This study was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia (Agreement no. 451-03-9/2021-14/200122 and 451-03-9/2021-14/200378), and these results are a part of Jovana V. Jovankić’s Ph.D. thesis.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest disclosure
The authors declare that they have no conflict of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Jovankić, J.V., Cvetković, D.M., Milutinović, M.G. et al. The impact of medicinal plant Ocimum minimum L. on fatty acid synthesis process in breast cancer cells. Biologia 77, 489–501 (2022). https://doi.org/10.1007/s11756-021-00939-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11756-021-00939-y