Prostat Kanserinde Yağ Damlacık Oluşumu ile İlişkili PLA2G7, UCP2 ve NEDD4L Genlerinin Protein Ekspresyonlarının Araştırılması

Öz

Amaç: Prostat kanseri (PCa) düzensiz lipid metabolizması ile karakterize edilen bir kanserdir. PCa hücrelerinin; lipitleri lipid damlacıkları şeklinde depoladığı ayrıca proliferasyon ve büyümeyi kolaylaştırmak amacıyla membran sentezi için yapıtaşı olarak kullandığı gözlemlenmiştir. Bu çalışmada amacımız PC3 ve DU145 prostat kanseri hücre hatlarında lipid damlacık metabolizmasında görev alan PLA2G7, UCP2 ve NEDD4L proteinlerinin ekspresyon seviyelerini incelemektir. Materyal ve Metot: Prostat kanseri hücre hatları orta seviye metastatik DU145 ve ileri metastatik PC3 %10 FBS, %1 penisilin-streptomisin, %1 non-esansiyel aminoasit içeren yüksek glukoz DMEM besi yerinde çoğaltılmıştır. Daha sonra PLA2G7, UCP2 ve NEDD4L proteinlerinin ekspresyon düzeyleri ELISA testi ile incelenmiştir. Bulgular: Bu çalışmada, PC3 ve DU145 hücrelerinde PLA2G7 protein seviyeleri açısından anlamlı bir fark olmadığı (p>0,05) ancak UCP2 protein seviyelerinin PC3 hücrelerinde anlamlı derecede arttığı gösterilmiştir (p<0,05). Bunun aksine NEDD4L protein seviyeleri DU145 hücre hattında anlamlı derecede düşmüştür (p<0,05). Sonuç: Çalışmanın sonucunda, UCP2'nin prostat kanserinin ilerlemesinde rolü ve NEDD4L ile hücre proliferasyonu kontrolü ile arasında bir ilişki olabilir.

Anahtar Kelimeler

• Lipid damlacığı
• NEDD4L
• PLA2G7
• Prostat kanseri
• UCP2

Investigation of Protein Expressions of PLA2G7, UCP2 and NEDD4L Genes Associated with Fat Droplet Formation in Prostate Cancer

Abstract

Objective: Prostate cancer (PCa) is characterised by dysregulated lipid metabolism. PCa cells store lipids as lipid droplets and use them to facilitate proliferation and growth. In this study, we aim to investigate the expression levels of PLA2G7, UCP2 and NEDD4L proteins, which are involved in lipid droplet metabolism, in PC3 (advanced metastatic) and DU145 (intermediate metastatic) PCa cells. Materials and Methods: DU145 and PC3 cells were cultured in a high-glucose DMEM medium containing 10% FBS, 1% penicillin-streptomycin, and 1% non-essential amino acid. The expression levels of PLA2G7, UCP2, and NEDD4L protein were assessed with ELISA assay. Results: There was no significant difference in the protein level of PLA2G7 between the PC3 and DU145 cells (p>0.05), while the protein level of UCP2 increased in the PC3 cell line significantly (p<0.05). The protein level of NEDD4L decreased significantly in the DU145 cell line when compared to the PC3 cell line (p<0.05). Conclusions: As a result of this study, the UCP2 gene might play a role in the progression of prostate cancer, and there could be a relationship between NEDD4L and cell proliferation control.

Keywords

• Lipid droplet
• NEDD4L
• PLA2G7
• Prostate cancer
• UCP2

References

1. Stoykova GE, Schlaepfer IR. Lipid metabolism and endocrine resistance in prostate cancer, and new opportunities for therapy. Int J Mol Sci. 2019;20(11):2626. doi:10.3390/ijms20112626
2. Jin C, Yuan P. Implications of lipid droplets in lung cancer: Associations with drug resistance. Oncol Lett. 2020;20(3):2091-2104. doi:10.3892/ol.2020.11769
3. Petan T, Jarc E, Jusović M. Lipid droplets in cancer: guardians of fat in a stressful world. Molecules. 2018;23(8):1941. doi:10.3390/molecules23081941
4. Schilke RM, Blackburn CMR, Bamgbose TT, Woolard MD. Interface of phospholipase activity, ımmune cell function, and atherosclerosis. Biomolecules. 2020;10(10):1449 doi:10.3390/biom10101449
5. Bermúdez MA, Balboa MA, Balsinde J. Lipid droplets, phospholipase A2, arachidonic acid, and atherosclerosis. Biomedicines. 2021;9(12):1891. Published 2021 Dec 13. doi:10.3390/biomedicines9121891
6. Luby A, Alves-Guerra MC. UCP2 is a Cancer target through energy metabolism and oxidative stress control. Int J Mol Sci. 2022;23(23):15077. doi:10.3390/ijms232315077
7. Diano S, Horvath TL. Mitochondrial uncoupling protein 2 (UCP2) in glucose and lipid metabolism. Trends Mol Med. 2012;18(1):52-58. doi:10.1016/j.molmed.2011.08.003
8. Xie S, Xia L, Song Y, Liu H, Wang ZW, Zhu X. Insights Into the biological role of NEDD4L E3 ubiquitin ligase in human cancers. Front Oncol. 2021;11:774648. doi:10.3389/fonc.2021.774648

9. Alberts P, Rotin D. Regulation of lipid droplet turnover by ubiquitin ligases. BMC Biol. 2010;8:94. doi:10.1186/1741-7007-8-94
10. Cruz ALS, Barreto EA, Fazolini NPB, Viola JPB, Bozza PT. Lipid droplets: platforms with multiple functions in cancer hallmarks. Cell Death Dis. 2020;11(2):105. doi:10.1038/s41419-020-2297-3
11. Bian X, Liu R, Meng Y, Xing D, Xu D, Lu Z. Lipid metabolism and cancer. J Exp Med. 2021;218(1):e20201606. doi:10.1084/jem.20201606
12. Roman M, Wrobel TP, Panek A, Paluszkiewicz C, Kwiatek WM. Lipid droplets in prostate cancer cells and effect of irradiation studied by Raman microspectroscopy. Biochim Biophys Acta Mol Cell Biol Lipids. 2020;1865(9):158753. doi:10.1016/j.bbalip.2020.158753
13. Benli E, Bayrak A, Cirakoglu A, Bayrak T, Noyan T. Comparison of serum acetylhydrolase (PAF-AH) and paraoxonase 1 (PON1) values between prostate cancer patients and a control group. Kaohsiung J Med Sci. 2017;33(11):572-577. doi:10.1016/j.kjms.2017.06.016
14. Lehtinen L, Vainio P, Wikman H, et al. PLA2G7 associates with hormone receptor negativity in clinical breast cancer samples and regulates epithelial-mesenchymal transition in cultured breast cancer cells. J Pathol Clin Res. 2017;3(2):123-138. doi:10.1002/cjp2.69
15. Stafforini DM, Zimmerman GA. Unraveling the PAF-AH/Lp-PLA2 controversy. J Lipid Res. 2014;55(9):1811-1814. doi:10.1194/jlr.E052886
16. Vainio P, Lehtinen L, Mirtti T, et al. Phospholipase PLA2G7, associated with aggressive prostate cancer, promotes prostate cancer cell migration and invasion and is inhibited by statins. Oncotarget. 2011;2(12):1176-1190. doi:10.18632/oncotarget.397
17. Kispert SE, Marentette JO, McHowat J. Enhanced breast cancer cell adherence to the lung endothelium via PAF acetylhydrolase inhibition: a potential mechanism for enhanced metastasis in smokers. Am J Physiol Cell Physiol. 2014;307(10):C951-C956. doi:10.1152/ajpcell.00218.2014
18. Segalés J, Sánchez-Martín C, Pujol-Morcillo A, et al. Role of UCP2 in the energy metabolism of the cancer cell line A549. Int J Mol Sci. 2023;24(9):8123.
19. Pecqueur C, Bui T, Gelly C, et al. Uncoupling protein-2 controls proliferation by promoting fatty acid oxidation and limiting glycolysis-derived pyruvate utilization. FASEB J. 2008;22(1):9-18. doi:10.1096/fj.07-8945com
20. Esteves P, Pecqueur C, Ransy C, et al. Mitochondrial retrograde signaling mediated by UCP2 inhibits cancer cell proliferation and tumorigenesis. Cancer Res. 2014;74(14):3971-3982. doi:10.1158/0008-5472.CAN-13-3383
21. Burch TC, Rhim JS, Nyalwidhe JO. Mitochondria biogenesis and bioenergetics gene profiles in isogenic prostate cells with different malignant phenotypes. Biomed Res Int. 2016;2016:1785201. doi:10.1155/2016/1785201
22. Song F, Li JZ, Wu Y, Wu WY, Wang Y, Li G. Ubiquitinated ligation protein NEDD4L participates in MiR-30a-5p attenuated atherosclerosis by regulating macrophage polarization and lipid metabolism. Mol Ther Nucleic Acids. 2021;26:1303-1317. doi:10.1016/j.omtn.2021.10.030
23. Hooper C, Puttamadappa SS, Loring Z, Shekhtman A, Bakowska JC. Spartin activates atrophin-1-interacting protein 4 (AIP4) E3 ubiquitin ligase and promotes ubiquitination of adipophilin on lipid droplets. BMC Biol. 2010;8:72. doi:10.1186/1741-7007-8-72
24. Hu XY, Xu YM, Fu Q, Yu JJ, Huang J. NEDD4L expression is downregulated in prostate cancer compared to benign prostatic hyperplasia. Eur J Surg Oncol. 2009;35(5):527- 531. doi:10.1016/j.ejso.2008.09.015