Specific binding of D-Amino neuraminic acid to ganglioside studied in prostate cancer cells

Kenan Demir; Hüseyin Aktuğ; Gürkan Yiğittürk; Eda Açıkgöz; Günnur Güler; Hadi Rouhrazi

TR EN

Prostat kanseri hücreleri'nde D-Amino nöraminik asidin gangliozid'e spesifik bağlanmasının çalışılması

Öz

Amaç: Bu çalışmanın amacı, insan D-Amino Nöraminik Asid’inin (KDN, 2-keto-3-deoksi-D-glisero-Dgalakto-nononik asit) hücresel bağlanma bölgesini araştırmaktır. KDN molekülü, sialik asit ailesinin bir üyesidir ve kanser hücrelerinde ekspresyonu artar. KDN'nin alabalık sperminde Monosialodihexosyl Gangliosid’e (GM3) bağlandığı gösterilmiştir. Gereç ve Yöntem: Bu çalışmada bir prostat kanseri hücre dizisi (DU145) kullanıldı. Her deney grubu; Kontrol, Glukosilseramid sentaz (GCS) enzim inhibitörü Genz-123346 ile tedavi edilen ve GM3 sentez inhibitörü Triptolid ile tedavi edilen olmak üzere 3 gruba ayrıldı. Her grup, GM3, Disialosyllactosylceramide (GD3) ve KDN için immünositokimyasal yöntem kullanılarak boyandı. Tedaviden sonra hücresel değişikliklerin sağlaması Fourier Transform Infrared (FTIR) Spektroskopi analizi ile yapıldı. Bulgular: Tedavi edilmeyen 1 numaralı hücre grubu, GM3, GD3 ve KDN ile pozitif boyandı ve GCS enzimi, sadece KDN ile pozitif boyanan 2 numaralı hücrelerin Genz-123346 grubuyla bloke edildi. Ayrıca, GD3 sentaz inhibitörü Triptolide ile muamele edilmiş 3 numaralı hücre grubu, GM3 ve KDN ile pozitif boyandı. FTIR ölçümleri Triptolide ile apoptotik özellikler gösterirken, Genz-123346 hücre canlılığı üzerinde olumsuz bir etkiye sahip değildi. Şeker yapılarında azalma ortaya çıktı ve immunositokimyasal boyama ile elde ettiğimiz sonuçlar FTIR ile pekiştirildi. Sonuç: KDN'nin yerinin belirlenmesi, kanser tedavisi araştırmaları için yeni hedeflerin seçilmesi açısından önemlidir. KDN'nin GM3 inhibisyonu ve GD3 inhibisyonu tarafından inhibe edilmediği gösterilmiştir. KDN, GM3 üzerinde olabileceği gibi farklı yerlere de bağlanabilir veya serbest halde olabilir. Bu çalışmada yalnızca salt GM veya GD serisindeki gangliozidlerden herhangi birine bağlanmayacağı ortaya konulmuştur.

Anahtar Kelimeler

Specific binding of D-Amino neuraminic acid to ganglioside studied in prostate cancer cells

Abstract

Aim: The aim of this study was to investigate the cellular binding site of human D-Amino Neuraminic Acid (KDN, 2-keto-3-deoxy-D-glycero-D-galacto-nononic acid). The KDN molecule is a member of the sialic acid family, and its expression increases in cancer cells. KDN has been shown to bind to Monosialodihexosyl Ganglioside (GM3) in trout sperm. Materials and Methods: In this study, a prostate cancer cell line (DU145) was used. Each experimental group was divided into 3 groups: Control, Glucosylceramide synthase (GCS) enzyme inhibitor Genz-123346-treated, and GM3 synthesis inhibitor Triptolide-treated. Each group was stained using the immunocytochemical method for GM3, Disialosyllactosylceramide (GD3) and KDN. Fourier Transform Infrared (FTIR) Spectroscopy analysis was performed to elucidate the cellular changes after treatment. Results: The group of non-treated number 1 cells stained positive with GM3, GD3 and KDN, and the GCS enzyme was blocked with the Genz-123346 group of number 2 cells stained positive only with KDN. Furthermore, the group of GD3 synthase inhibitor Triptolide treated number 3 cells stained positive with GM3 and KDN. FTIR measurements showed apoptotic characteristics with Triptolide, while Genz-123346 did not have a negative effect on cell viability. There was a reduction in sugar structures and the results obtained with immunocytochemical staining were reinforced with FTIR. Conclusions: Determining the location of the bound KDN is important for the selection of new targets for cancer treatment research. KDN has been shown to be not inhibited by GM3 inhibition and GD3 inhibition. KDN can be on GM3 as well as connected to different places or can be free. In this study, it was demonstrated that it would not bind to any of the gangliosides in the pure GM or GD series.

Keywords

Kaynakça

Battula VL, Shi Y, Evans KW, Wang RY, Spaeth EL, Jacamo RO, et al. Ganglioside GD2 identifies breast cancer stem cells and promotes tumorigenesis. J Clin Invest. 2012;122(6):2066-78. https://doi.org/10.1172/JCI59735
Sarkar TR, Battula VL, Werden SJ, Vijay GV, Ramirez-Peña EQ, Taube JH, et al. GD3 synthase regulates epithelial-mesenchymal transition and metastasis in breast cancer. Oncogene. 2015;34(23):2958-67. https://doi.org/10.1038/onc.2014.245
Ponnapakam AP, Liu J, Bhinge KN, Drew BA, Wang TL, Antoon JW, et al. 3-Ketone-4,6-diene ceramide analogs exclusively induce apoptosis in chemo-resistant cancer cells. Bioorg Med Chem. 2014;22(4):1412-20. https://doi.org/10.1016/j.bmc.2013.12.065
Chung TW, Choi HJ, Kim SJ, Kwak CH, Song KH, Jin UH, et al. The ganglioside GM3 is associated with cisplatin-induced apoptosis in human colon cancer cells. PLoS One. 2014;9(5):e92786. https://doi.org/10.1371/journal.pone.0092786
Lehninger AL, David LN, Cox MM. Lipids. In: Lehninger AL, David LN (ed) Lehninger Principles of Biochemistry, New York: W.H. 2005:357. https://doi: 10.1007/978-3-662-08289-8
Ledeen RW, Wu G. GM1 ganglioside: another nuclear lipid that modulates nuclear calcium. GM1 potentiates the nuclear sodium-calcium exchanger. Can J Physiol Pharmacol. 2006;84(3-4):393-402. https://doi.org/10.1139/y05-133
Utz JR, Crutcher T, Schneider J, Sorgen P, Whitley CB. Biomarkers of central nervous system inflammation in infantile and juvenile gangliosidoses. Mol Genet Metab. 2015;114(2):274-80. https://doi.org/10.1016/j.ymgme.2014.11.015
Krengel U, Bousquet PA. Molecular recognition of gangliosides and their potential for cancer immunotherapies. Front Immunol. 2014;21:325. https://doi.org/10.3389/fimmu.2014.00325

Liu YY, Hill RA, Li YT. Ceramide glycosylation catalyzed by glucosylceramide synthase and cancer drug resistance. Adv Cancer Res. 2013;117:59-89. https://doi.org/10.1016/B978-0-12-394274-6.00003-0
Nadano D, Iwasaki M, Endo S, Kitajima K, Inoue S, Inoue YA. Naturally occurring deaminated neuraminic acid, 3-deoxy-D-glycero-D-galacto-nonulosonic acid (KDN). Its unique occurrence at the nonreducing ends of oligosialyl chains in polysialoglycoprotein of rainbow trout eggs. J Biol Chem. 1986;261(25):11550-7.
Yu S, Kojima N, Hakomori SI, Kudo S, Inoue S, Inoue Y. Binding of rainbow trout sperm to egg is mediated by strong carbohydrate-to-carbohydrate interaction between (KDN)GM3 (deaminated neuraminyl ganglioside) and Gg3-like epitope. Proc Nat Acad Sci USA. 2002;99(5):2854-9. https://doi.org/10.1073/pnas.052707599
Güler G, Acikgoz E, Karabay Yavasoglu NÜ, Bakan B, Goormaghtigh E, Aktug H. Deciphering the biochemical similarities and differences among mouse embryonic stem cells, somatic and cancer cells using ATR-FTIR spectroscopy. Analyst. 2018;143(7):1624-34. https://doi.org/10.1039/c8an00017d
Acikgoz E, Güler G, Camlar M, Oktem G, Aktug H. Glycogen synthase kinase-3 inhibition in glioblastoma multiforme cells induces apoptosis, cell cycle arrest and changing biomolecular structure. Spectrochim Acta A Mol Biomol Spectrosc. 2019;209:150-64. https://doi.org/10.1016/j.saa.2018.10.036
Güler G, Guven U, Oktem G. Characterization of CD133+/CD44+ human prostate cancer stem cells with ATRFTIR spectroscopy. Analyst. 2019;144(6):2138-49. https://doi.org/10.1039/C9AN00093C
Barth A. Infrared spectroscopy of proteins. BBA – Bioenergetics. 2007;1767(9):1073-101. https://doi.org/10.1016/j.bbabio.2007.06.004
Güler G, Gärtner RM, Ziegler C, Mäntele W. Lipid-Protein Interactions in the Regulated Betaine Symporter BetP Probed by Infrared Spectroscopy. J Biol Chem. 2016;291(9):4295-307. https://doi.org/10.1074/jbc.M114.621979
Karman J, Tedstone JL, Gumlaw NK, Hu Y, Yew N, Siegel C, et al. Reducing glycosphingolipid biosynthesis in airway cells partially ameliorates disease manifestations in a mouse model of asthma. Int Immunol. 2010;22(7):593-603. https://doi.org/10.1093/intimm/dxq044
Natoli TA, Smith LA, Rogers KA, Wang B, Komarnitsky S, Budman Y, et al. Inhibition of glucosylceramide accumulation results in effective blockade of polycystic kidney disease in mouse models. Nat Med. 2010;16(7):788-92. https://doi.org/10.1038/nm.2171
Kwon HY, Kim SJ, Kim CH, Son SW, Kim KS, Lee JH, et al. Triptolide downregulates human GD3 synthase (hST8Sia I) gene expression in SK-MEL-2 human melanoma cells. Exp Mol Med. 2010;42(12):849-55. https://doi.org/10.3858/emm.2010.42.12.088
Slevin M, Costello B, Kumar S, Gaffney J. The ganglioside GM(3) is raised in the sera and tissue of patients with bladder tumours. Int J Oncol. 1996;8(2):271-4. https://doi.org/10.3892/ijo.8.2.271
Kacuráková M, Mathlouthi M. FTIR and laser-Raman spectra of oligosaccharides in water: characterization of the glycosidic bond. Carbohydr Res. 1996;284(2):145-57. https://doi.org/10.1016/0008-6215(95)00412-2
Lewis AT, Jones K, Lewis KE, Jones S, Lewis PD. Detection of Lewis Antigen Structural Change by FTIR Spectroscopy. Carbohydr Polym. 2013;92(2):1294-301. https://doi.org/10.1016/j.carbpol.2012.09.078
Le Roux K, Prinsloo LC, Meyer D. Fourier Transform Infrared spectroscopy discloses different types of cell death in flow cytometrically sorted cells. Toxicol In Vitro. 2015;29(7):1932-40. https://doi.org/10.1016/j.tiv.2015.08.002
Plaimee P, Weerapreeyakul N, Thumanu K, Tanthanuch W, Barusrux S. Melatonin induces apoptosis through biomolecular changes, in SK-LU-1 human lung adenocarcinoma cells. Cell Prolif. 2014;47(6):564-77. https://doi.org/10.1111/cpr.12140
Gao Y, Huo X, Dong L, et al. Fourier transform infrared microspectroscopy monitoring of 5-fluorouracil-induced apoptosis in SW620 colon cancer cells. Mol Med Rep. 2015;11(4):2585-91. https://doi.org/10.3892/mmr.2014.3088.

Ayrıntılar

Birincil Dil

İngilizce

Konular

Histoloji ve Embriyoloji

Bölüm

Araştırma Makalesi

Yazarlar

Kenan Demir ^*
0000-0003-2864-6041
Türkiye

Hüseyin Aktuğ
0000-0003-4150-8495
Türkiye

Gürkan Yiğittürk
0000-0002-5315-253X
Türkiye

Eda Açıkgöz
0000-0002-6772-3081
Türkiye

Günnur Güler
0000-0002-8485-7372
Türkiye

Hadi Rouhrazi
0000-0002-1367-064X
Türkiye

Yayımlanma Tarihi

12 Haziran 2023

Gönderilme Tarihi

5 Ekim 2022

Kabul Tarihi

7 Aralık 2022

Yayımlandığı Sayı

Yıl 1970 Cilt: 62 Sayı: 2

IZ

https://izlik.org/JA68FP75SY

Kaynak Göster

RIS / Bibtex

Vancouver

1.Kenan Demir, Hüseyin Aktuğ, Gürkan Yiğittürk, Eda Açıkgöz, Günnur Güler, Hadi Rouhrazi. Specific binding of D-Amino neuraminic acid to ganglioside studied in prostate cancer cells. ETD [Internet]. 01 Haziran 2023;62(2):301-9. Erişim adresi: https://izlik.org/JA68FP75SY