Caffeic Acid Phenethyl Ester Alleviates Cryodamage To Lung Cancer Cells During Cryopreservation

Year 2021, , 247 - 253, 29.09.2021

Ezgi Avşar Abdik

https://doi.org/10.17350/HJSE19030000235

Abstract

Cryopreservation is widely used technique for long-term preservation of viable cells at low temperature. In this process, considering the effects of cryodamage on cells, the application of safe and efficient cryoprotective agents is very important. Caffeic acid phenethyl ester (CAPE) is a natural biological compound which is found in propolis extract and possess beneficial effects such as anti-oxidant, antimicrobial, anti-inflammatory. In the current study was to investigate the cryoprotective effects of CAPE on human lung cancer cell line, A549. Firstly, cells were cryopreserved in freezing medium with/without different concentrations of CAPE (5, 10, and 20 μM). The cells were frozen slowly and kept in liquid nitrogen for one month. After thawing, the cryoprotective effects of CAPE were determined by cell viability, proliferation, colony formation, and gene expression levels. The results showed that 5μM CAPE supplemented freezing medium significantly increased the viability of post thaw A549 cells. 5μM CAPE treatment significantly increased cell proliferation after 24, 48 and 72h since thawing compared to control. 10 μM CAPE did not significantly affect cell viability compared to control group. Also, 5μM CAPE increased the number of A549 colonies compared to 10μM CAPE and control groups. Furthermore, markedly larger colonies were noticed in 5μM CAPE group. In addition, 5μM CAPE significantly increased apoptosis and proliferation-related genes, Akt, NFκB and Bcl-2, expression levels compared to 10μM CAPE and control groups. CAPE may be a potential cryoprotective agent for relieving cryodamage during cryopreservation.

Keywords

Cryopreservation, Cryoprotective agents, CAPE, Lung cancer cell line, Cell Viability.

Supporting Institution

Yeditepe University

References

• 1. Pegg DE. Principles of cryopreservation. In: Cryopreservation and Freeze-Drying Protocols. Springer, pp. 3-19, 2015.
• 2. Gonda K, Shigeura T, Sato T, et al. Preserved proliferative capacity and multipotency of human adipose-derived stem cells after long-term cryopreservation. Plastic and reconstructive surgery 121(2) (2008) 401-410.
• 3. Nunes VA, Gozzo AJ, Cruz-Silva I, et al. Vitamin E prevents cell death induced by mild oxidative stress in chicken skeletal muscle cells. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 141(3) (2005) 225-240.
• 4. Joshi A. A review and application of cryoprotectant: The science of cryonics. PharmaTutor 4(1) (2016) 12-18.
• 5. Luzar A, Chandler D. Structure and hydrogen bond dynamics of water--dimethyl sulfoxide mixtures by computer simulations. The Journal of chemical physics 98(10) (1993) 8160-8173.
• 6. Hornberger K, Yu G, McKenna D, Hubel A. Cryopreservation of hematopoietic stem cells: emerging assays, cryoprotectant agents, and technology to improve outcomes. Transfusion Medicine and Hemotherapy 46(3) (2019) 188-196.
• 7. Borrelli F, Maffia P, Pinto L, et al. Phytochemical compounds involved in the anti-inflammatory effect of propolis extract. Fitoterapia 73 (2002) S53--S63.
• 8. Koltuksuz U, Irmak MK, Karaman A, et al. Testicular nitric oxide levels after unilateral testicular torsion/detorsion in rats pretreated with caffeic acid phenethyl ester. Urological research 28(6) (2000) 360-363.
• 9. Son S, Lewis BA. Free radical scavenging and antioxidative activity of caffeic acid amide and ester analogues: Structure- activity relationship. Journal of agricultural and food chemistry 50(3) (2002) 468-472.
• 10. Zheng Z, Yenari MA. Post-ischemic inflammation: molecular mechanisms and therapeutic implications. Neurological research 26(8) (2004) 884-892.
• 11. Guney M, Oral B, Karahan N, Mungan T. Protective effect of caffeic acid phenethyl ester (CAPE) on fluoride-induced oxidative stress and apoptosis in rat endometrium. Environmental toxicology and pharmacology 24(2) (2007) 86-91.
• 12. Parlakpinar H, Sahna E, Acet A, Mizrak B, Polat AI. Protective effect of caffeic acid phenethyl ester (CAPE) on myocardial ischemia--reperfusion-induced apoptotic cell death. Toxicology 209(1) (2005) 1-14.
• 13. Amodio R, De Ruvo C, Di Matteo V, et al. Caffeic acid phenethyl ester blocks apoptosis induced by low potassium in cerebellar granule cells. International journal of developmental neuroscience 21(7) (2003) 379-389.
• 14. Isildar B, Ozkan S, Oncul M, et al. Comparison of different cryopreservation protocols for human umbilical cord tissue as source of mesenchymal stem cells. Acta histochemica 121(3) (2019) 361-367.
• 15. Ninagawa T, Eguchi A, Kawamura Y, Konishi T, Narumi A. A study on ice crystal formation behavior at intracellular freezing of plant cells using a high-speed camera. Cryobiology 73(1) (2016) 20-29.
• 16. Mazur P. Kinetics of water loss from cells at subzero temperatures and the likelihood of intracellular freezing. The Journal of general physiology 47(2) (1963) 347-369.
• 17. Matsumura K, Hyon S-H. Polyampholytes as low toxic efficient cryoprotective agents with antifreeze protein properties. Biomaterials 30(27) (2009) 4842-4849.
• 18. Aliakbari F, Gilani MAS, Amidi F, et al. Improving the efficacy of cryopreservation of spermatogonia stem cells by antioxidant supplements. Cellular Reprogramming (Formerly" Cloning and Stem Cells") 18(2) (2016) 87-95.
• 19. Shaban S, El-Husseny MWA, Abushouk AI, Salem AMA, Mamdouh M, Abdel-Daim MM. Effects of antioxidant supplements on the survival and differentiation of stem cells. Oxidative Medicine and Cellular Longevity 2017 (2017).
• 20. Bugger H, Pfeil K. Mitochondrial ROS in myocardial ischemia reperfusion and remodeling. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease 1866(7) (2020) 165768.
• 21. Singh R, Cuervo AM. Autophagy in the cellular energetic balance. Cell metabolism 13(5) (2011) 495-504.
• 22. Feng T-Y, Li Q, Ren F, et al. Melatonin Protects Goat Spermatogonial Stem Cells against Oxidative Damage during Cryopreservation by Improving Antioxidant Capacity and Inhibiting Mitochondrial Apoptosis Pathway. Oxidative Medicine and Cellular Longevity 2020 (2020).
• 23. Song YS, Park E-H, Hur GM, et al. Caffeic acid phenethyl ester inhibits nitric oxide synthase gene expression and enzyme activity. Cancer letters 175(1) (2002) 53-61.
• 24. Mahdi NS, Azarbani F, Pirnia A, Abbaszadeh A, Gholami M. The Effect of Caffeic Acid on Spermatogonial Stem Cell-type A Cryopreservation. Reports of Biochemistry & Molecular Biology 7(1) (2018) 85.
• 25. Soleimanzadeh A, Talavi N, Yourdshahi VS, Bucak MN. Caffeic acid improves microscopic sperm parameters and antioxidant status of buffalo (Bubalus bubalis) bull semen following freeze-thawing process. Cryobiology 95 (2020) 29-35.
• 26. da Silva CM, Macias-Garcia B, Miró-Morán A, et al. Melatonin reduces lipid peroxidation and apoptotic-like changes in stallion spermatozoa. Journal of pineal research 51(2) (2011) 172-179.
• 27. Liu Y, Zhang B, Zhang J, et al. CAPE promotes the expansion of human umbilical cord blood-derived hematopoietic stem and progenitor cells in vitro. Science China Life Sciences 57(2) (2014) 188-194.
• 28. Çalışkan M, Pritchard JK, Ober C, Gilad Y. The effect of freeze-thaw cycles on gene expression levels in lymphoblastoid cell lines. PloS one 9(9) (2014) e107166.
• 29. Koppers AJ, Mitchell LA, Wang P, Lin M, Aitken RJ. Phosphoinositide 3-kinase signalling pathway involvement in a truncated apoptotic cascade associated with motility loss and oxidative DNA damage in human spermatozoa. Biochemical Journal 436(3) (2011) 687-698.
• 30. Downward J. PI 3-kinase, Akt and cell survival. In: Seminars in Cell & Developmental Biology. Vol 15. ; 2004:177-182.
• 31. Najafi A, Adutwum E, Yari A, et al. Melatonin affects membrane integrity, intracellular reactive oxygen species, caspase3 activity and AKT phosphorylation in frozen thawed human sperm. Cell and tissue research 372(1) (2018) 149-159.
• 32. Fahy BN, Schlieman M, Virudachalam S, Bold RJ. AKT inhibition is associated with chemosensitisation in the pancreatic cancer cell line MIA-PaCa-2. British journal of cancer 89(2) (2003) 391-397.
• 33. Braun F, de Carné Trécesson S, Bertin-Ciftci J, Juin P. Protect and serve: Bcl-2 proteins as guardians and rulers of cancer cell survival. Cell Cycle 12(18) (2013) 2937-2947.
• 34. Uz E, Söğüt S, Şahin Ş, et al. The protective role of caffeic acid phenethyl ester (CAPE) on testicular tissue after testicular torsion and detorsion. World journal of urology 20(4) (2002) 264-270.
• 35. Dilber Y, Inan S, Ercan GA, Sencan A. The role of CAPE in PI3K/AKT/mTOR activation and oxidative stress on testis torsion. Acta histochemica 118(1) (2016) 31-37.
• 36. Tolba MF, Omar HA, Azab SS, Khalifa AE, Abdel-Naim AB, Abdel-Rahman SZ. Caffeic acid phenethyl ester: a review of its antioxidant activity, protective effects against ischemia-reperfusion injury and drug adverse reactions. Critical reviews in food science and nutrition 56(13) (2016) 2183-2190.