Effect of whey protein derivatives on cell viability, cell migration and cell cycle phases in MCF-7 cells

Year 2023, , 39 - 45, 31.01.2023

F. Tutku Aksoy Ayse Mine Yılmaz Gokhan Bıcım A. Suha Yalcın

https://doi.org/10.5472/marumj.1244676

Abstract

Objective: This study aimed to obtain protein derivatives after treatment of whey proteins with hazelnut oil and olive oil and determined
their effects on MCF-7 cells.
Materials and Methods: Whey proteins obtained from 6% whey powder were treated with hazelnut oil (HO) and olive oil (OO) at a
protein to lipid ratio of 1:10 at 60 ̊C for 120 minutes. The protein derivatives formed with whey protein and HO or OO were applied
to MCF-7 cancer cells and healthy fibroblasts. The effects of protein derivatives on cell viability, apoptosis, reactive oxygen species
(ROS) production, wound healing, cell cycle phase distribution and cell cycle related proteins Akt and p21(Waf1/Cip1) expressions
were investigated.
Results: Cell viability decreased significantly after 24 h of incubation with WP:OO. The percentage of apoptotic or necrotic cells varied
between 5-10% and no statistically significant effect was observed. There was no statistically significant difference in ROS production
and colony formation between controls and WP:HO or WP:OO groups. Treatment of cells with WP:OO for 24 h significantly decreased
cell migration compared to the control group. G2/M phase was significantly suppressed in WP:OO group compared to the control
group. WP:OO also increased the expression of p21(Waf1/Cip1) significantly when compared with the control group.
Conclusion: Our results showed that whey protein derivatives applied to MCF-7 cells are cytotoxic and may be useful in breast cancer
treatment.

Keywords

Whey proteins, Oleic acid, Cell survival, Apoptosis, Cell cycle

References

• Ensminger AH, Ensminger ME, Konlande JE, Robson JRK. Milk and milk products. The Concise Encylopedia of Foods and Nutrition. CRC Press Inc USA 1995;691-710.
• Bounous G. Whey protein concentrate (WPC) and glutathione modulation in cancer treatment. Anticancer Res 2000;20:6:4785-92.
• De Wit JN. Nutritional and functional characteristics of whey proteins in food products. J Dairy Sci 1998;81:3:597-608. doi. org/10.3168/jds.S0022-0302(98)75613-9
• Smithers GW. Whey and whey proteins-From “gutter-togold.” Int Dairy J 2008;18:7:695–704. doi.org/10.1016/j. idairyj.2008.03.008
• Smithers GW, Ballard FJ, Copeland AD, et al. New opportunities from the isolation and utilization of whey proteins. J Dairy Sci 1996;79:8:1454-9. doi.org/10.3168/jds. S0022-0302(96)76504-9
• Yalçın AS. Emerging therapeutic potential of whey proteins and peptides. Curr Pharm Des 2006;12:13:1637-43. doi.org/10 .2174/138.161.206776843296
• Düringer C, Hamiche A, Gustafsson L, Kimura H, Svanborg C. HAMLET interacts with histones and chromatin in tumor cell nuclei. J Biol Chem 2003;278:43:42131-42135. doi. org/10.1074/jbc.M306462200
• Mok KH, Pettersson J, Orrenius S, Svanborg C. HAMLET, protein folding, and tumor cell death. Biochem Biophys Res Commun 2007;354:1:1-7. doi.org/10.1016/j.bbrc.2006.12.167
• Svensson M, Fast J, Mossberg AK, et al. Alpha-lactalbumin unfolding is not sufficient to cause apoptosis, but is required for the conversion to HAMLET (human alpha-lactalbumin made lethal to tumor cells). Protein Sci 2003;12:2794-804. doi. org/10.1110/ps.0231003
• Pettersson J, Mossberg AK, Svanborg C. α-Lactalbumin species variation, HAMLET formation, and tumor cell death. Biochem Biophys Res Commun 2006;345:1:260-70. doi. org/10.1016/j.bbrc.2006.04.081
• Svensson M, Håkansson A, Mossberg A, Linse S, Svanborg C. Conversion of alpha-lactalbumin to a protein inducing apoptosis. Proc Natl Acad Sci U S A 2000;97:8:4221-26. doi. org/10.1073/pnas.97.8.4221
• Fischer W, Gustafsson L, Mossberg AK, et al. Human α-lactalbumin made lethal to tumor cells (HAMLET) kills human glioblastoma cells in brain xenografts by an apoptosis-like mechanism and prolongs survival. Cancer Res 2004;64:6:2105-12. doi.org/10.1158/0008-5472.CAN-03-2661
• Mossberg AK, Wullt B, Gustafsson L, Månsson W, Ljunggren E, Svanborg C. Bladder cancers respond to intravesical instillation of HAMLET (human α-lactalbumin made lethal to tumor cells). Int J Cancer 2007;121:6:1352-9. doi.org/10.1002/ ijc.22810
• Fang B, Zhang M, Tian M, Jiang L, Guo HY, Ren FZ. Bovine lactoferrin binds oleic acid to form an anti-tumor complex similar to HAMLET. Biochim Biophys Acta Mol Cell Biol Lipids 2014;1841:4:535-43. doi.org/10.1016/j.bbalip.2013.12.008
• Brinkmann CR, Thiel S, Otzen DE. Protein-fatty acid complexes: Biochemistry, biophysics and function. FEBS J 2013;280:8:1733-49. doi.org/10.1111/febs.12204
• Fontana A, Spolaore B, de Laureto PP. The biological activities of protein/oleic acid complexes reside in the fatty acid. Biochim Biophys Acta Proteins Proteom 2013;1834:6:1125-43. doi. org/10.1016/j.bbapap.2013.02.041
• Permyakov SE, Knyazeva EL, Khasanova LM, et al. Oleic acid is a key cytotoxic component of HAMLET-like complexes. Biol Chem 2012;393:1-2:85-92. doi.org/10.1515/BC-2011-230
• Farràs M, Almanza-Aguilera E, Hernáez Á, et al. Beneficial effects of olive oil and Mediterranean diet on cancer physiopathology and incidence. Semin Cancer Biol 2021;73:178-95. doi.org/10.1016/j.semcancer.2020.11.011
• Casaburi I, Puoci F, Chimento A, et al. Potential of olive oil phenols as chemopreventive and therapeutic agents against cancer: a review of in vitro studies. Mol Nutr Food Res 2013;57:1:71-83. doi.org/10.1002/mnfr.201200503
• Borzì AM, Biondi A, Basile F, Luca S, Vicari ESD, Vacante M. Olive oil effects on colorectal cancer. Nutrients 2018;11:1:32. doi.org/10.3390/nu11010032
• Rodríguez-Miguel C, Moral R, Escrich R, Vela E, Solanas M, Escrich E. The role of dietary extra virgin olive oil and corn oil on the alteration of epigenetic patterns in the rat DMBA-induced breast cancer model. PLoS One 2015;10:9: e0138980. doi.org/10.1371/journal.pone.0138980
• Corominas-Faja B, Cuyàs E, Lozano-Sánchez J, et al. Extravirgin olive oil contains a metabolo-epigenetic inhibitor of cancer stem cells. Carcinogenesis 2018;39:4:601-13. doi. org/10.1093/carcin/bgy023
• Colomer R, Menéndez JA. Mediterranean diet, olive oil and cancer. Clin Transl Oncol 2006;8:1:15-21. doi.org/10.1007/ s12094.006.0090-0
• Reboredo-Rodríguez P. Varela-López A, Forbes-Hernández TY, et al. Phenolic compounds isolated from olive oil as nutraceutical tools for the prevention and management of cancer and cardiovascular diseases. Int J Mol Sci 2018;19:8:2305. doi.org/10.3390/ijms19082305
• Chen Z, Zhang B, Gao F, Shi R. Modulation of G2/M cell cycle arrest and apoptosis by luteolin in human colon cancer cells and xenografts. Oncol Lett 2018;15:2:1559-65. doi. org/10.3892/ol.2017.7475
• Benitez-Sánchez PL, León-Camacho M, Aparicio R. A comprehensive study of hazelnut oil composition with comparisons to other vegetable oils, particularly olive oil. Eur Food Res Technol 2003;218:1:13-9. doi.org/10.1007/ s00217.003.0766-4
• Bayram T, Pekmez M, Arda N, Yalçın AS. Antioxidant activity of whey protein fractions isolated by gel exclusion chromatography and protease treatment. Talanta 2008;75:3:705-9. doi. org/10.1016/j.talanta.2007.12.007
• Brinkmann CR, Brodkorb A, Thiel S, Kehoe JJ. The cytotoxicity of fatty acid/α‐lactalbumin complexes depends on the amount and type of fatty acid. Eur J Lipid Sci Technol 2013;115:6:591- 600. doi.org/10.1002/ejlt.201200165
• Liu J, Ke F, Xu Z, et al. CCR6 is a prognostic marker for overall survival in patients with colorectal cancer, and its overexpression enhances metastasis in vivo. PLoS One 2014;9:6:e101137. doi.org/10.1371/journal.pone.0101137
• Zhang M, Yang F, Yang F, Chen J, Zheng CY, Liang Y. Cytotoxic aggregates of α-lactalbumin induced by unsaturated fatty acid induce apoptosis in tumor cells. Chem Biol Interact 2009;180:2:131-42. doi.org/10.1016/j.cbi.2009.03.019
• Brinkmann CR, Thiel S, Larsen MK, Petersen TE, Jensenius JC, Heegaard CW. Preparation and comparison of cytotoxic complexes formed between oleic acid and either bovine or human α-lactalbumin. J Dairy Sci 2011;94:5:2159-70. doi. org/10.3168/jds.2010-3622
• Trulsson M, Yu H, Gisselsson L, et al. HAMLET binding to α-actinin facilitates tumor cell detachment. PLoS One 2011;6:3:e.17179. doi.org/10.1371/journal.pone.0017179
• Yao Q, Li H, Fan L, Huang S, Wang J, Zheng N. The combination of lactoferrin and linolenic acid inhibits colorectal tumor growth through activating AMPK/JNK-related apoptosis pathway. PeerJ 2021;9:e11072. doi.org/10.7717/peerj.11072
• El-Fakharany EM, Abu-Serie MM, Litus EA, et al. The use of human, bovine, and camel milk albumins in anticancer complexes with oleic acid. Protein J 2018;37:3:203-15. doi. org/10.1007/s10930.018.9770-1
• Lišková K, Auty MAE, Chaurin V, et al. Cytotoxic complexes of sodium oleate with β-lactoglobulin. Eur J Lipid Sci Technol 2011;113:10:1207-18. doi.org/10.1002/ejlt.201100109
• Brinkmann CR, Heegaard CW, Petersen TE, Jensenius JC, Thiel S. The toxicity of bovine α-lactalbumin made lethal to tumor cells is highly dependent on oleic acid and induces killing in cancer cell lines and noncancer-derived primary cells. FEBS J 2011;278:11:1955-967. doi.org/10.1111/j.1742- 4658.2011.08112.x