Modulation of sphingolipid metabolism: Investigation of the apoptotic and cytotoxic effects of the ceramidase Inhibitor D-e-MAPP in HT29 colon cancer cells

Abstract

Purpose: Colon cancer is one of the most common cancers of the digestive system and usually develops when polyps develop into cancer. Factors such as age, genetic predisposition and dietary habits increase the risk of the disease. Therefore, early diagnosis of colon cancer and development of appropriate clinical approaches are of great importance today. Sphingolipids are essential biological molecules that serve as important components of cell membranes and play important roles in various cellular processes. They are involved in various basic biological processes such as cell growth and differentiation, especially apoptosis (programmed cell death) and intracellular signaling. Sphingolipids are divided into different subgroups such as ceramide, sphingomyelin and glycosphingolipid and coordinate the design and activities of cells and play important roles in anticancer processes such as cell cycle arrest and inducing apoptosis. Dysfunction of sphingolipid metabolism plays a critical role in the development and progression of numerous cancers. Altered sphingolipid balance in cancer cells can affect the regulation of processes such as cellular growth, invasion and metastasis. In particular, changes in ceramide levels can activate cell survival pathways and cause cancer to become more aggressive. Method: In order to investigate the potential anticancer effects of ceramide inhibitor, we investigated the effects of D-e MAPP, a ceramidase inhibitor, on HT29 colon cancer cell line. For this purpose, cell viability and morphological changes after treatment were evaluated in detail. Cell viability was determined by MTT assay to determine the cytotoxic effects of treatment. Furthermore, structural and morphological changes in cells were visualized by confocal microscopy and treatment-induced cell death modes were determined by flow cytometry to obtain detailed information on cell death pathways. Findings: These methods aim to contribute to a more comprehensive understanding of the effects of ceramidase inhibitors on colon cancer cells. Conclusion: This study aims to contribute to a more comprehensive understanding of the effects of ceramidase inhibitors such as D-e-MAPP on colon cancer cells. By assessing cell viability, morphological changes and cell death pathways, this research provides valuable insights into the potential therapeutic applications of targeting sphingolipid metabolism in colon cancer treatment. Keywords: Colon cancer, sphingolipids, D-e-MAPP

Keywords

• Colon cancer
• sphingolipids
• D-e-MAPP

Sfingolipid Metabolizmasının Modülasyonu: Seramidaz İnhibitörü D-e-MAPP'ın HT29 Kolon Kanseri Hücrelerinde Apoptotik ve Sitotoksik Etkilerinin Araştırılması

Abstract

Amaç: Kolon kanseri en sık görülen sindirim sistemi kanserlerinden biridir ve genellikle poliplerin kansere dönüşmesiyle gelişir. Yaş, genetik yatkınlık ve beslenme alışkanlıkları gibi faktörler hastalık riskini artırmaktadır. Bu nedenle kolon kanserinin erken teşhisi ve uygun klinik yaklaşımların geliştirilmesi günümüzde büyük önem taşımaktadır. Sfingolipidler, *Corresponding author Tel.: [phone]; E-mail:[e-mail] © Copyright 2025 by Biological Diversity and Conservation Geliş tarihi: 19.12.2024; Yayın tarihi: 15.12.2025 BioDiCon. 1184-191224 284 Biological Diversity and Conservation – 18 / 3 (2025) hücre membranlarının önemli bileşenleri olarak görev yapan ve çeşitli hücresel süreçlerde önemli roller oynayan temel biyolojik moleküllerdir. Hücre büyümesi ve farklılaşması, özellikle apoptoz (programlanmış hücre ölümü) ve hücre içi sinyalizasyon gibi çeşitli temel biyolojik süreçlere katılırlar. Sfingolipidler seramid, sfingomiyelin ve glikosfingolipid gibi farklı alt gruplara ayrılır ve hücrelerin tasarımını ve faaliyetlerini koordine eder ve hücre döngüsünü durdurma ve apoptozu indükleme gibi antikanser süreçlerinde önemli roller oynar. Sfingolipid metabolizması işlev bozukluğu, çok sayıda kanserin gelişimi ve ilerlemesinde kritik bir rol oynamaktadır. Kanser hücrelerinde sfingolipid dengesinin değişmesi hücresel büyüme, invazyon ve metastaz gibi süreçlerin düzenlenmesini etkileyebilir. Özellikle, seramid seviyelerindeki değişiklikler hücre hayatta kalma yollarını aktive edebilir ve kanserin daha agresif hale gelmesine neden olabilir. Metod:Seramid inhibitörünün potansiyel antikanser etkilerini araştırmak amacıyla, bu çalışmada bir seramidaz inhibitörü olan D-e-MAPP'nin HT29 kolon kanseri hücre hattı üzerindeki etkileri araştırılmıştır. Bu amaçla, tedavi sonrası hücre canlılığı ve morfolojik değişiklikler ayrıntılı olarak değerlendirilmiştir. Hücre canlılığı, tedavinin sitotoksik etkilerini belirlemek için MTT testi ile belirlenmiştir. Ayrıca, hücrelerdeki yapısal ve morfolojik değişiklikler konfokal mikroskopi ile görüntülenmiş ve hücre ölüm yolları hakkında ayrıntılı bilgi edinmek için tedaviye bağlı hücre ölüm modları akış sitometrisi ile belirlenmiştir. Bulgular: Bu yöntemler, seramidaz inhibitörlerinin kolon kanseri hücreleri üzerindeki etkilerinin daha kapsamlı bir şekilde anlaşılmasına katkıda bulunmayı amaçlamaktadır. Sonuç: Bu çalışma, D-e-MAPP gibi seramidaz inhibitörünün kolon kanseri hücreleri üzerindeki etkilerinin daha kapsamlı bir şekilde anlaşılmasına katkıda bulunmayı amaçlamaktadır. Hücre canlılığını, morfolojik değişiklikleri ve hücre ölüm yollarını değerlendiren bu araştırma, kolon kanseri tedavisinde sfingolipid metabolizmasını hedeflemenin potansiyel terapötik uygulamaları hakkında değerli bilgiler sunmaktadır.

Keywords

• Kolon kanseri
• sfingolipidler
• D-e-MAPP

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