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

References

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Details

Primary Language

English

Subjects

Histology and Embryology

Journal Section

Research Article

Authors

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

Publication Date

June 12, 2023

Submission Date

October 5, 2022

Acceptance Date

December 7, 2022

Published in Issue

Year 1970 Volume: 62 Number: 2

IZ

https://izlik.org/JA68FP75SY

Cite

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. EJM [Internet]. 2023 Jun. 1;62(2):301-9. Available from: https://izlik.org/JA68FP75SY