Alginate nanoparticles containing Rosmarinus officinalis essential oil and α-pinene: cytotoxicity and effect on apoptotic-involved genes in human melanoma and breast cancer cell lines

Document Type : Original Research Article

Authors

1 Department of Biochemistry, School of Medicine, Fasa University of Medical Sciences, Fasa, Iran

2 Department of Medical Genetics, School of Medicine, Fasa University of Medical Sciences, Fasa, Iran

3 Estahban Higher Education Center- Shiraz University, Estahban, Iran

4 Department of Tissue Engineering and Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran

5 Noncommunicable Diseases Research Center, Fasa University of Medical Sciences, Fasa, Iran

6 Department of medical nanotechnology, schools of advanced technologies in medicine, Fasa university of medical sciences (FUMS), Fasa, Iran.

Abstract

Cancers are one of the major causes of death, and the development of new medicine using advanced technologies such as nanotechnology has thus received more attention. Rosmarinus officinalis as a common medicinal plant possess many biological effects, such as anticancer effects. This study investigated the chemical compositions of its essential oil using GC-MS analysis. Alpha-pinene (24.0%), eucalyptol (16.99%), verbenone (8.79%), L-borneol (7.79%), and camphor (6.87%) were the five major compounds. After that, alginate nanoparticles containing α-pinene (major compound) and R. officinalis essential oil were prepared with particle sizes of 137 ± 8 and 151 ± 7. Besides, successful loading of α-pinene or the essential oil in nanoparticles was confirmed using ATR-FTIR analysis. The efficacy (IC50) of alginate nanoparticles containing α-pinene against A-375 and MCF-7 were 582 (179-1886) µg/mL and 94 (47-187) µg/mL. These values for alginate nanoparticles containing R. officinalis essential oil were obtained as 807 (470-1383) µg/mL and 235 (168-328) µg/mL. Moreover, RT-PCR results show that the Bcl2 (anti-apoptotic gene) level in MCF-7 treated cells with alginate nanoparticles containing R. officinalis essential oil was higher than Bax (apoptotic gene); another cell death pathway rather than apoptosis was involved. Considering the proper efficacy of alginate nanoparticles containing the essential oil or α-pinene, especially against MCF-7 cells, they could be considered for further investigation in vivo.

Keywords

Main Subjects


  1. WHO. Cancer Ket Facts. Available at: https://www.who.int/news-room/fact-sheets/detail/cancer. Accessed Agust, 2022.
  2. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians, 2018;68 (6):394-424.https://doi.org/10.3322/caac.21492
  3. Toma A-O, Prodan M, Reddyreddy AR, Seclaman E, Crainiceanu Z, Bloanca V, Bratosin F, Dumitru C, Pilut CN, Alambaram S, Vasamsetti NG, Decean L, Pricop M. The Epidemiology of Malignant Melanoma during the First Two Years of the COVID-19 Pandemic: A Systematic Review. International Journal of Environmental Research and Public Health, 2023;20 (1):305.https://doi.org/10.3390/ijerph20010305
  4. Domingues B, Lopes JM, Soares P, Pópulo H. Melanoma treatment in review. ImmunoTargets and therapy, 2018;7:35.https://doi.org/10.2147/ITT.S134842
  5. Davis LE, Shalin SC, Tackett AJ. Current state of melanoma diagnosis and treatment. Cancer Biology & Therapy, 2019;20 (11):1366-1379.https://doi.org/10.1080/15384047.2019.1640032
  6. Rebecca VW, Sondak VK, Smalley KS. A brief history of melanoma: from mummies to mutations. Melanoma Research, 2012;22 (2):114-122.https://doi.org/10.1097/CMR.0b013e328351fa4d
  7. Nieto G, Huvaere K, Skibsted LH. Antioxidant activity of rosemary and thyme by-products and synergism with added antioxidant in a liposome system. European Food Research and Technology, 2011;233 (1):11-18.https://doi.org/10.1007/s00217-011-1486-9
  8. Noorpisheh Ghadimi S, Sharifi N, Osanloo M. The leishmanicidal activity of essential oils: A systematic review. Journal of Herbmed Pharmacology, 2020;9 (4):300-308.https://doi.org/10.34172/jhp.2020.38
  9. Comşa Ş, Cimpean AM, Raica M. The story of MCF-7 breast cancer cell line: 40 years of experience in research. Anticancer Research, 2015;35 (6):3147-3154. 
  10. Cavo M, Fato M, Peñuela L, Beltrame F, Raiteri R, Scaglione S. Microenvironment complexity and matrix stiffness regulate breast cancer cell activity in a 3D in vitro model. Scientific Reports, 2016;6 (1):1-13.https://doi.org/10.1038/srep35367
  11. Wu S, Wang Y, Yuan Z, Wang S, Du H, Liu X, Wang Q, Zhu X. Human adipose‑derived mesenchymal stem cells promote breast cancer MCF7 cell epithelial‑mesenchymal transition by cross interacting with the TGF‑β/Smad and PI3K/AKT signaling pathways. Molecular Medicine Reports, 2019;19 (1):177-186.https://doi.org/10.3892/mmr.2018.9664
  12. Fayyad RJ, Ali ANM, Dwaish AS, Al-Abboodi AKA. Anticancer activity of Spirulina platensis methanolic extracts against L20B and MCF7 human cancer cell lines. Plant Arch, 2019;19 (1):1419-1426. 
  13. Massi D, Brusa D, Merelli B, Ciano M, Audrito V, Serra S, Buonincontri R, Baroni G, Nassini R, Minocci D. PD-L1 marks a subset of melanomas with a shorter overall survival and distinct genetic and morphological characteristics. Annals of Oncology, 2014;25 (12):2433-2442.https://doi.org/10.1093/annonc/mdu452
  14. Vincent KM, Postovit L-M. Investigating the utility of human melanoma cell lines as tumour models. Oncotarget, 2017;8 (6):10498.https://doi.org/10.18632/oncotarget.14443
  15. Yu M-H, Choi J-H, Chae I-G, Im H-G, Yang S-A, More K, Lee I-S, Lee J. Suppression of LPS-induced inflammatory activities by Rosmarinus officinalis L. Food Chemistry, 2013;136 (2):1047-1054.https://doi.org/10.1016/j.foodchem.2012.08.085
  16. Nieto G, Ros G, Castillo J. Antioxidant and antimicrobial properties of rosemary (Rosmarinus officinalis, L.): A review. Medicines, 2018;5 (3):98.https://doi.org/10.3390/medicines5030098
  17. Ngo SN, Williams DB, Head RJ. Rosemary and cancer prevention: preclinical perspectives. Critical Reviews in Food Science and Nutrition, 2011;51 (10):946-954.https://doi.org/10.1080/10408398.2010.490883
  18. Park BB, An JY, Park SU. Recent studies on pinene and its biological and pharmacological activities. EXCLI journal, 2021;20:812-818. 
  19. Allegra A, Tonacci A, Pioggia G, Musolino C, Gangemi S. Anticancer activity of Rosmarinus officinalis L.: mechanisms of action and therapeutic potentials. Nutrients, 2020;12 (6):1739.https://doi.org/10.3390/nu12061739
  20. Kelidari HR, Alipanah H, Roozitalab G, Ebrahimi M, Osanloo M. Anticancer Effect of Solid-Lipid Nanoparticles Containing Mentha longifolia and Mentha pulegium Essential Oils: In Vitro Study on Human Melanoma and Breast Cancer Cell Lines. Biointerface Research in Applied Chemistry, 2021;12 (2):2128-2137.https://doi.org/10.33263/BRIAC122.21282137
  21. Qasemi H, Fereidouni Z, Karimi J, Abdollahi A, Zarenezhad E, Rasti F, Osanloo M. Promising antibacterial effect of impregnated nanofiber mats with a green nanogel against clinical and standard strains of Pseudomonas aeruginosa and Staphylococcus aureus. J Drug Deliv Sci Technol, 2021;66:102844.https://doi.org/10.1016/j.jddst.2021.102844
  22. Fernando IPS, Lee W, Han EJ, Ahn G. Alginate-based nanomaterials: Fabrication techniques, properties, and applications. Chemical Engineering Journal, 2020;391:123823.https://doi.org/10.1016/j.cej.2019.123823
  23. Zeng Y, Xiang Y, Sheng R, Tomás H, Rodrigues J, Gu Z, Zhang H, Gong Q, Luo K. Polysaccharide-based nanomedicines for cancer immunotherapy: A review. Bioactive Materials, 2021;6 (10):3358-3382.https://doi.org/10.1016/j.bioactmat.2021.03.008
  24. Hariyadi DM, Islam N. Current Status of Alginate in Drug Delivery. Adv Pharmacol Pharm Sci, 2020;2020:8886095.https://doi.org/10.1155/2020/8886095
  25. Moemenbellah-Fard MD, Shahriari-Namadi M, Kelidari HR, Nejad ZB, Ghasemi H, Osanloo M. Chemical composition and repellent activity of nine medicinal essential oils against Anopheles stephensi, the main malaria vector. Int J Trop Insect Sci, 2021;41 (2):1325-1332.https://doi.org/10.1007/s42690-020-00325-2
  26. Sanei-Dehkordi A, Moemenbellah-Fard MD, Saffari M, Zarenezhad E, Osanloo M. Nanoliposomes containing limonene and limonene-rich essential oils as novel larvicides against malaria and filariasis mosquito vectors. BMC Complementary Medicine and Therapies, 2022;22 (1):140.https://doi.org/10.1186/s12906-022-03624-y
  27. Hamidpour R, Hamidpour S, Elias G. Rosmarinus officinalis (Rosemary): a novel therapeutic agent for antioxidant, antimicrobial, anticancer, antidiabetic, antidepressant, neuroprotective, anti-inflammatory, and anti-obesity treatment. Biomed J Sci Tech Res, 2017;1 (4):1-6.https://doi.org/10.26717/BJSTR.2017.01.000371
  28. Kamyar K, Hiva A, Hadi B, Negar R, Mahmoud O. Chitosan Nanoparticles Containing Cinnamomum verum J.Presl Essential Oil and Cinnamaldehyde: Preparation, Characterization and Anticancer Effects against Melanoma and Breast Cancer Cells. Traditional and Integrative Medicine, 2022;7 (1). 
  29. Alipanah H, Rasti F, Zarenezhad E, Dehghan A, Sahebnazar B, Osanloo M. Comparison of Anticancer Effects of Carvone, Carvone-Rich Essential Oils, and Chitosan Nanoparticles Containing Each of Them. Biointerface Res Appl Chem, 2022;12:5716-5726.https://doi.org/10.33263/BRIAC124.57165726
  30. Valizadeh A, Khaleghi AA, Alipanah H, Zarenezhad E, Osanloo M. Anticarcinogenic Effect of Chitosan Nanoparticles Containing Syzygium aromaticum Essential Oil or Eugenol Toward Breast and Skin Cancer Cell Lines. BioNanoScience, 2021.https://doi.org/10.1007/s12668-021-00880-z
  31. Alipanah H, Farjam M, Zarenezhad E, Roozitalab G, Osanloo M. Chitosan nanoparticles containing limonene and limonene-rich essential oils: potential phytotherapy agents for the treatment of melanoma and breast cancers. BMC Complementary Medicine and Therapies, 2021;21 (1):1-10.https://doi.org/10.1186/s12906-021-03362-7
  32. Rogers C, Erkes DA, Nardone A, Aplin AE, Fernandes-Alnemri T, Alnemri ES. Gasdermin pores permeabilize mitochondria to augment caspase-3 activation during apoptosis and inflammasome activation. Nature communications, 2019;10 (1):1-17.https://doi.org/10.1038/s41467-019-09397-2
  33. Nagata S. Apoptosis and clearance of apoptotic cells. Annual Review of Immunology, 2018;36:489-517.https://doi.org/10.1146/annurev-immunol-042617-053010
  34. Kulsoom B, Shamsi TS, Afsar NA, Memon Z, Ahmed N, Hasnain SN. Bax, Bcl-2, and Bax/Bcl-2 as prognostic markers in acute myeloid leukemia: are we ready for Bcl-2-directed therapy? Cancer Management and Research, 2018;10:403-416.https://doi.org/10.2147/CMAR.S154608
  35. Pistritto G, Trisciuoglio D, Ceci C, Garufi A, D'Orazi G. Apoptosis as anticancer mechanism: function and dysfunction of its modulators and targeted therapeutic strategies. Aging, 2016;8 (4):603-619.https://doi.org/10.18632/aging.100934
  36. Xu Q, Li M, Yang M, Yang J, Xie J, Lu X, Wang F, Chen W. α-pinene regulates miR-221 and induces G2/M phase cell cycle arrest in human hepatocellular carcinoma cells. Bioscience Reports, 2018;38 (6).https://doi.org/10.1042/BSR20180980
  37. Matsuo AL, Figueiredo CR, Arruda DC, Pereira FV, Scutti JAB, Massaoka MH, Travassos LR, Sartorelli P, Lago JH. α-Pinene isolated from Schinus terebinthifolius Raddi (Anacardiaceae) induces apoptosis and confers antimetastatic protection in a melanoma model. Biochemical and Biophysical Research Communications, 2011;411 (2):449-454.https://doi.org/10.1016/j.bbrc.2011.06.176
  38. D'Arcy MS. Cell death: a review of the major forms of apoptosis, necrosis and autophagy. Cell Biology International, 2019;43 (6):582-592.https://doi.org/10.1002/cbin.11137
  39. Pérez-Sánchez A, Barrajón-Catalán E, Ruiz-Torres V, Agulló-Chazarra L, Herranz-López M, Valdés A, Cifuentes A, Micol V. Rosemary (Rosmarinus officinalis) extract causes ROS-induced necrotic cell death and inhibits tumor growth in vivo. Scientific Reports, 2019;9 (1):1-11.https://doi.org/10.1038/s41598-018-37173-7
  40. Golstein P, Kroemer G. Cell death by necrosis: towards a molecular definition. Trends in Biochemical Sciences, 2007;32 (1):37-43.https://doi.org/10.1016/j.tibs.2006.11.001