Öz
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.
Anahtar Kelimeler
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.
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
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 Makaleleri |
| Yazarlar | |
| Yayımlanma Tarihi | 12 Haziran 2023 |
| Gönderilme Tarihi | 5 Ekim 2022 |
| Yayımlandığı Sayı | Yıl 2023Cilt: 62 Sayı: 2 |
