The Effects of Rosemary (Rosmarinus Officinalis L.) Extract on the Proliferation and Apoptosis of A549 and H1299 Human Lung Cancer Cells

Year 2024, , 293 - 300, 27.10.2024

Ebru Alimoğulları , Bahar Kartal , Tuba Özdemir Sancı , Sinem Aslan Erdem , Aslı Ceylan

https://doi.org/10.53446/actamednicomedia.1474338

Abstract

ABSTRACT
Objective: Apoptosis resistance and increased proliferation rates are characteristics of cancer cells. The anticancer properties of rosemary (Rosmarinus officinalis L.) extract (RE) have been demonstrated in a small number of in vivo and in vitro animal studies; however, no research has investigated the role of RE in human non-small cell lung cancer (NSCLC) A549 and H1299 cells, and its underlying mechanism of action remains unknown. In the current study, we examined the effects of RE on human non-small cell lung cancer cell proliferation, survival, and apoptosis.
Methods: Human non-small cell lung cancer (NSCLC) A549 and H1299 were incubated with (2.5 µg/ml, 5 µg/ml, 7.5 µg/ml, 10 µg/ml, and 12.5 µg/ml) doses of RE for 12, 24, and 48 hours. MTT, Annexin V-PI, and caspase 3/7 assay kit were performed to detect the cell viability and apoptosis.
Results: According to MTT analysis, the viability of A549 and H1299 human lung cancer cells was reduced by approximately 49.74% and 47.76%, respectively, for 24 hours by treatment with a dose of 5 µg/ml RE. The results of Annexin V-PI staining and Caspase 3/7 activation showed that RE had a greater effect on inducing cell death.
Conclusion: In conclusion, we can say that rosemary extract has both apoptotic and antiproliferative properties on human lung cancer cells. We might propose that additional investigation is necessary to ascertain the therapeutic impacts of rosemary extract.

Keywords

Annexin V-PI, A549, Caspase 3/7, H1299, Rosemary extract.

Biberiye (Rosmarinus Officinalis L.) Ekstresinin A549 ve H1299 İnsan Akciğer Kanseri Hücrelerinin Çoğalması ve Apoptozu Üzerindeki Etkileri

Abstract

ÖZ
Amaç: Apoptoz direnci ve artan proliferasyon oranları kanser hücrelerinin karakteristik özelliğidir. Biberiye (Rosmarinus officinalis L.) ekstresinin (RE) antikanser özellikleri, az sayıda in vivo ve in vitro hayvan çalışmasında gösterilmiştir; ancak RE'nin insandaki Küçük Hücreli Olmayan Akciğer Kanseri (NSCLC) A549 ve H1299 hücrelerindeki rolünü araştıran araştırma yoktur ve bunun altında yatan etki mekanizması belirsizliğini korumaktadır. Bu çalışmada RE'nin insandaki Küçük Hücreli Olmayan Akciğer Kanseri hücre hatlarının çoğalması, canlılığı ve apoptoz üzerindeki etkilerini araştırdık.
Yöntem: İnsandaki Küçük Hücreli Olmayan Akciğer Kanseri (NSCLC) A549 ve H1299 hücre hatları 12, 24 ve 48 saat süre boyunca (2.5 µg/ml, 5 µg/ml, 7.5 µg/ml, 10 µg/ml ve 12.5 µg/ml) RE dozlarıyla inkübe edildi. Hücre canlılığı ve apoptoz belirlemek için MTT, Annexin V-PI ve kaspaz 3/7 kiti kullanıldı.
Bulgular: MTT analizine göre, A549 ve H1299 insan akciğer kanseri hücrelerinin canlılığı, 5 µg/ml RE doz tedavisiyle 24 saat boyunca yaklaşık sırasıyla % 49,74 ve % 47,76 oranında azaldı. Annexin V-PI ve Kaspaz 3/7 aktivasyonunun sonuçları, RE'nin hücre ölümünü indüklemede büyük bir etkiye sahip olduğunu gösterdi.
Sonuç: Sonuç olarak biberiye ekstresinin insan akciğer kanseri hücreleri üzerinde hem apoptotik hem de antiproliferatif özelliklere sahip olduğunu söyleyebiliriz. Biberiye ekstresinin terapötik etkilerini belirlemek için daha fazla araştırmaya ihtiyaç vardır.

Keywords

Annexin V-PI, A549, Biberiye ekstresi, Caspase 3/7, H1299

References

• Stewart BW, Wild CP. World Cancer Report 2014. Lyon, France. International Agency for Research. 2014. Reck M, Rabe KF. Precision Diagnosis and Treatment for Advanced Non-Small-Cell Lung Cancer. N Engl J Med. 2017;377(9):849-861. doi:10.1056/NEJMra1703413
• Pao W, Chmielecki J. Rational, biologically based treatment of EGFR-mutant non-small-cell lung cancer. Nat Rev Cancer. 2010;10(11):760-774. doi:10.1038/nrc2947
• Araujo L.H., Horn L., Merritt R.E., Shilo K., Meng X.-W., Carbone D.P. Cancer of the Lung: Non-Small Cell Lung Cancer and Small Cell Lung Cancer. Abeloff’s Clinical Oncology. Elsevier; Philadelphia, PA, USA: 2019; pp. 1108-1158.
• Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011 Mar 4;144(5):646-74. doi:10.1016/j.cell.2011.02.013
• da Rocha AB, Lopes RM, Schwartsmann G. Natural products in anticancer therapy. Curr Opin Pharmacol. 2001;1(4):364-369. doi:10.1016/s1471-4892(01)00063-7.
• Cuvelier ME, Berset C, Richard H. Antioxidant Constituents in Sage (Salvia officinalis). J. Agric. Food Chem. 1994; 42, 665–669. doi:10.1021/jf00039a012
• Pertino MW, Theoduloz C, Butassi E, Zacchino S, Schmeda-Hirschmann G. Synthesis, antiproliferative and antifungal activities of 1,2,3-triazole-substituted carnosic acid and carnosol derivatives. Molecules. 2015;14;20(5):8666-86. doi:10.3390/molecules20058666
• Bernardes WA, Lucarini R, Tozatti MG, Souza MG, Silva ML, Filho AA, Martins CH, Crotti AE, Pauletti PM, Groppo M, Cunha WR. Antimicrobial activity of Rosmarinus officinalis against oral pathogens: relevance of carnosic acid and carnosol. Chem Biodivers. 2010;7(7):1835-40. doi:10.1002/cbdv.200900301
• Yesil-Celiktas O, Sevimli C, Bedir E, Vardar-Sukan F. Inhibitory effects of rosemary extracts, carnosic acid and rosmarinic acid on the growth of various human cancer cell lines. Plant Foods Hum Nutr. 2010;65(2):158-63. doi:10.1007/s11130-010-0166-4
• Kelsey NA, Wilkins HM, Linseman DA. Nutraceutical antioxidants as novel neuroprotective agents. Molecules. 2010;15(11):7792-814. doi:10.3390/molecules15117792
• Lopez-Jimenez A., Garcia-Caballero M., Medina M.A., Quesada A.R. Antiangiogenic properties of carnosol and carnosic acid, two major dietary compounds from rosemary. Eur. J. Nutr. 2013; 52: 85–95. doi:10.1007/s00394-011-0289-x
• Bauer J., Kuehnl S., Rollinger J.M., Scherer O., Northoff H., Stuppner H. et al. Carnosol and carnosic acids from Salvia officinalis inhibit microsomal prostaglandin E2 synthase-1. J. Pharmacol. Exp. Ther.2012; 342: 169–176. doi:10.1124/jpet.112.193847
• Costa S., Utan A., Speroni E., Cervellati R., Piva G., Prandini A., et al. Carnosic acid from rosemary extracts: a potential chemoprotective agent against aflatoxin B1. An in vitro study. J. Appl. Toxicol. 2007; 27: 152–159. doi:10.1002/jat.1186
• Osakabe N, Takano H, Sanbongi C, Yasuda A, Yanagisawa R, Inoue K, Yoshikawa T. Anti-inflammatory and anti-allergic effect of rosmarinic acid (RA); inhibition of seasonal allergic rhinoconjunctivitis (SAR) and its mechanism. Biofactors. 2004;21(1-4):127-31. doi:10.1002/biof.552210125
• Qiao S, Li W, Tsubouchi R, Haneda M, Murakami K, Takeuchi F, Nisimoto Y, Yoshino M. Rosmarinic acid inhibits the formation of reactive oxygen and nitrogen species in RAW264.7 macrophages. Free Radic Res. 2005;39(9):995-1003. doi:10.1080/10715760500231836
• Sui X., Liu T., Ma C., Yang L., Zu Y., Zhang L., et al. Microwave irradiation to pretreat rosemary (Rosmarinus officinalis L.) for maintaining antioxidant content during storage and to extract essential oil simultaneously. Food. Chem. 2012; 131: 1399–1405. doi:10.1016/j.foodchem.2011.10.007
• Kim DS, Kim HR, Woo ER, Hong ST, Chae HJ, Chae SW. Inhibitory effects of rosmarinic acid on adriamycin-induced apoptosis in H9c2 cardiac muscle cells by inhibiting reactive oxygen species and the activations of c-Jun N-terminal kinase and extracellular signal-regulated kinase. Biochem Pharmacol. 2005;70(7):1066-78. doi:10.1016/j.bcp.2005.06.026
• Li GS, Jiang WL, Tian JW, Qu GW, Zhu HB, Fu FH. In vitro and in vivo antifibrotic effects of rosmarinic acid on experimental liver fibrosis. Phytomedicine. 2010;17(3-4):282-8. doi:10.1016/j.phymed.2009.05.002
• Fallarini S, Miglio G, Paoletti T, Minassi A, Amoruso A, Bardelli C, Brunelleschi S, Lombardi G. Clovamide and rosmarinic acid induce neuroprotective effects in in vitro models of neuronal death. Br J Pharmacol. 2009;157(6):1072-84. doi:10.1111/j.1476-5381.2009.00213.x
• Moore, Jessy, Mark Megaly, Adam J. MacNeil, Panagiota Klentrou and Evangelia Litsa Tsiani. Rosemary extract reduces Akt/mTOR/p70S6K activation and inhibits proliferation and survival of A549 human lung cancer cells. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 83 2016; 725-732. doi:10.1016/j.biopha.2016.07.043
• Cheung S., Tai J. Anti-proliferative and antioxidant properties of rosemary Rosmarinus officinalis. Oncol. Rep. 2007;17:1525–1531. doi:10.3892/or.17.6.1525
• González-Vallinas M, Molina S, Vicente G, Sánchez-Martínez R, Vargas T, García-Risco MR, Fornari T, Reglero G, Ramírez de Molina A. Modulation of estrogen and epidermal growth factor receptors by rosemary extract in breast cancer cells. Electrophoresis. 2014;35(11):1719-27. doi:10.1002/elps.201400011
• Yesil-Celiktas O, Sevimli C, Bedir E, Vardar-Sukan F. Inhibitory effects of rosemary extracts, carnosic acid and rosmarinic acid on the growth of various human cancer cell lines. Plant Foods Hum Nutr. 2010;65(2):158-63. doi:10.1007/s11130-010-0166-4
• Kontogianni VG, Tomic G, Nikolic I, Nerantzaki AA, Sayyad N, Stosic-Grujicic S, Stojanovic I, Gerothanassis IP, Tzakos AG. Phytochemical profile of Rosmarinus officinalis and Salvia officinalis extracts and correlation to their antioxidant and anti-proliferative activity. Food Chem. 2013;136(1):120-9. doi:10.1016/j.foodchem.2012.07.091
• Petiwala SM, Berhe S, Li G, Puthenveetil AG, Rahman O, Nonn L, Johnson JJ. Rosemary (Rosmarinus officinalis) extract modulates CHOP/GADD153 to promote androgen receptor degradation and decreases xenograft tumor growth. PLoS One. 2014;9(3):e89772. doi:10.1371/journal.pone.0089772
• Vicente G., Molina S., Gonzalez-Vallinas M., Garcia-Risco M.R., Fornari T., Reglero G., De Molina A.R. Supercritical rosemary extracts, their antioxidant activity and effect on hepatic tumor progression. Journal of Supercritical Fluids 2013; 79, pp. 101-108. doi:10.1016/j.supflu.2012.07.006
• Sancheti G, Goyal PK. Effect of Rosmarinus officinalis in modulating 7,12-dimethylbenz(a)anthracene induced skin tumorigenesis in mice. Phytother Res. 2006;20(11):981-6. doi:10.1002/ptr.1989
• Sancheti G, Goyal P. Modulatory influence of Rosemarinus officinalis on DMBA-induced mouse skin tumorigenesis. Asian Pac J Cancer Prev. 2006;7(2):331-5. PMID: 16839234.
• González-Vallinas M, Molina S, Vicente G, Zarza V, Martín-Hernández R, García-Risco MR, Fornari T, Reglero G, Ramírez de Molina A. Expression of microRNA-15b and the glycosyltransferase GCNT3 correlates with antitumor efficacy of rosemary diterpenes in colon and pancreatic cancer. PLoS One. 2014;9(6):e98556. doi:10.1371/journal. pone.0098556.
• Yan M, Li G, Petiwala SM, Householter E, Johnson JJ. Standardized rosemary (Rosmarinus officinalis) extract induces Nrf2/sestrin-2 pathway in colon cancer cells. Journal of Functional Foods. 2015;13:137-147. doi:10.1016/j.jff.2014.12.038
• Moore J, Megaly M, MacNeil AJ, Klentrou P, Tsiani E. Rosemary extract reduces Akt/mTOR/p70S6K activation and inhibits proliferation and survival of A549 human lung cancer cells. Biomed Pharmacother. 2016;83:725-732. doi:10.1016/j.biopha.2016.07.043. Epub 2016 Jul 29. PMID: 27470574.
• Newman DJ, Cragg GM. Natural products as sources of new drugs over the last 25 years. J Nat Prod. 2007;70(3):461-77. doi:10.1021/np068054v
• Kim HK, Wilson EG, Choi YH, Verpoorte R. Metabolomics: a tool for anticancer lead-finding from natural products. Planta Med. 2010;76(11):1094-102. doi:10.1055/s-0030-1249898
• Rahmani AH, Al Shabrmi FM, Allemailem KS, Aly SM, Khan MA. Implications of green tea and its constituents in the prevention of cancer via the modulation of cell signalling pathway. Biomed Res Int. 2015;2015:925640. doi:10.1155/2015/925640
• Moore J, Yousef M, Tsiani E. Anticancer Effects of Rosemary (Rosmarinus officinalis L.) Extract and Rosemary Extract Polyphenols. Nutrients. 2016;8(11):731. doi:10.3390/nu8110731
• Massi A, Bortolini O, Ragno D, Bernardi T, Sacchetti G, Tacchini M, De Risi C. Research Progress in the Modification of Quercetin Leading to Anticancer Agents. Molecules. 2017;22(8):1270. doi:10.3390/molecules22081270
• Shamshoum H, Vlavcheski F, Tsiani E. Anticancer effects of oleuropein. Biofactors. 2017;43(4):517-528. doi:10.1002/biof.1366
• Valdés A, Cifuentes A, León C. Foodomics evaluation of bioactive compounds in foods. TrAC Trends Anal Chem 2017; 96:2-13. doi:10.1016/j.trac.2017.06.004
• Sedighi R, Zhao Y, Yerke A, Sang S. Preventive and protective properties of rosemary (Rosmarinus officinalis L.) in obesity and diabetes mellitus of metabolic disorders: a brief review. Curr Opin Food Sci 2015:58–70. doi:10.1016/j.cofs.2015.02.002
• Karadağ AE, Demirci B, Çaşkurlu A, Demirci F, Okur ME, Orak D, Sipahi H, Başer KHC. In vitro antibacterial, antioxidant, anti-inflammatory and analgesic evaluation of Rosmarinus officinalis L. flower extract fractions. S Afr J Bot 2019;125:214–220. doi:10.1016/j.sajb.2019.07.039
• Tai J, Cheung S, Wu M, Hasman D. Antiproliferation effect of Rosemary (Rosmarinus officinalis) on human ovarian cancer cells in vitro. Phytomedicine. 2012;19(5):436-43. doi:10.1016/j.phymed.2011.12.012
• Urquiza-López A, Álvarez-Rivera G, Ballesteros-Vivas D, Cifuentes A, Del Villar-Martínez AA. Metabolite Profiling of Rosemary Cell Lines with Antiproliferative Potential against human HT-29 Colon Cancer Cells. Plant Foods Hum Nutr. 2021;76(3):319-325. doi:10.1007/s11130-021-00892-w
• Jaglanian A, Termini D, Tsiani E. Rosemary (Rosmarinus officinalis L.) extract inhibits prostate cancer cell proliferation and survival by targeting Akt and mTOR. Biomed Pharmacother. 2020. doi:10.1016/j.biopha.2020.110717
• Jaglanian A, Tsiani E. Rosemary Extract Inhibits Proliferation, Survival, Akt, and mTOR Signaling in Triple-Negative Breast Cancer Cells. Int J Mol Sci. 2020;21(3):810. doi:10.3390/ijms21030810
• Cattaneo L, Cicconi R, Mignogna G, Giorgi A, Mattei M, Graziani G, et al. Anti- Proliferative Effect of Rosmarinus officinalis L. Extract on Human Melanoma A375 cells. PLoS ONE 2015; 10(7): e0132439. doi:10.1371/journal.pone.0132439
• Gao Q., Liu H., Yao Y., Geng L., Zhang X., Jiang L., et al. Carnosic acid induces autophagic cell death through inhibition of the Akt/mTOR pathway in human hepatoma cells. J.Appl. Toxicol.2015; 35: 485– 492. doi:10.1002/jat.3049
• Levine CB, Bayle J, Biourge V, Wakshlag JJ. Cellular effects of a turmeric root and rosemary leaf extract on canine neoplastic cell lines. BMC Vet Res. 2017;13(1):388. doi:10.1186/s12917-017-1302-2
• O'Neill EJ, Moore J, Song J, Tsiani EL. Inhibition of non-small cell lung cancer proliferation and survival by rosemary extract is associated with activation of ERK and AMPK. Life (Basel). 2021;12(1):52. Published 2021 Dec 31. doi:10.3390/life12010052
• Gîrd CE, Costea T, Mitran V. Evaluation of cytotoxic activity and anticancer potential of indigenous Rosemary (Rosmarinus officinalis L.) and oregano (Origanum vulgare L.) dry extracts on MG-63 bone osteosarcoma human cell line. Rom J Morphol Embryol. 2021;62(2):525-535. doi:10.47162/RJME.62.2.19
• Makaremi S, Mosayebi G, Ghazavi A, Ganji A. Combination effect of ginger and rosemary extracts on ct-26 mouse colorectal cancer cells. Int J Basic Sci Med. 2022;7(3):114-120. doi:10.34172/ijbsm.2022.21
• Jang YG, Hwang KA, Choi KC. Rosmarinic Acid, a Component of Rosemary Tea, Induced the Cell Cycle Arrest and Apoptosis through Modulation of HDAC2 Expression in Prostate Cancer Cell Lines. Nutrients. 2018;10(11):1784. doi:10.3390/nu10111784