Investigation of D-e-MAPP-derived Cytotoxicity on Human Prostate Cancer Cells

Yıl 2019, Cilt: 6, 204 - 212, 30.09.2019

Mehtap Kutlu , Hüseyin İzgördü , Emre Çömlekçi Canan Vejselova Sezer

Öz

Sphingolipids play critical role in
biological processes such cell death, survival and drug resistance in many
cancers. Ceramide and dihydroceramide are proliferation and death associated
sphingolipids. Recent cancer research are focused on clarifying
cancer-sphingolipid metabolism. In last years, ceramide as a key molecule in
sphingolipid metabolism relationship has been investigated for its anticancer
activity via augmenting its intracellular level by ceramidase inhibitor
application. Prostate cancer is amoung the most frequent human cancers and is
reported as second most common cancer-related death cause. Prostate cancer is
common at ages older than 65. The aim of this study was to investigate the
potential cytotoxic activity of a ceramidease inhibitor
(1S,2R)-D-erythro-2-(N-Myristoylamino)-1-phenyl-1-propanol (D-erythro-MAPP) on
human prostate cancer DU-145 cells by using MTT
(3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay, flow
cytometry, confocal and TEM microscopy. MTT findings showed that D-erythro-MAPP
caused toxicity in low doses on DU-145 cells. Confocal and TEM microscopy
findings indicated hole formation in cytoskeleton, chromatin condensation,
horseshoe-shaped cell nuclei as morphological changes and blebbings on cell
membrane, fragmentation of nuclei, chromatin condensation and loss of criastae
of as ultrastructural changes, respectively. Flow cytometry findings showed
that D-erythro-MAPP triggered apoptosis in short term application of 24 hours
on DU-145 cells. According to results it can be conculded that D-erythro-MAPP
decreased viability of DU-145 cells in dose-dependent manner.  Our findings stated the anti-cancer and
cytotoxic potential of D-erythro-MAPP on DU-145 and this agent might be used in
drug developing for cancer treatment after the further in vitro and in vivo
studies.

Anahtar Kelimeler

Sphingolipid, DU-145, Confocal microscopy

İnsan Prostat Kanseri Hücrelerinde D-e-MAPP kaynaklı Sitotoksisitenin Araştırılması

Öz

Sfingolipidler,
birçok kanserde hücre ölümü, hayatta kalma ve ilaç direnci gibi biyolojik
işlemlerde kritik rol oynamaktadır. Seramid ve dihidroseramid, proliferasyon ve
ölüme bağlı olan sfingolipidlerdir. Son kanser araştırmaları,
kanser-sfingolipid metabolizmasının netleştirilmesine odaklanmıştır. Son
yıllarda, sfingolipid metabolizması ilişkisinde kilit bir molekül olarak
seramid, antikanser aktivitesi için seramidaz inhibitör uygulaması ile hücre
içi seviyesini arttırarak araştırılmıştır. Prostat kanseri, en sık görülen
insan kanserleri arasındadır ve kansere bağlı en yaygın ikinci ölüm nedeni
olarak rapor edilmektedir. Prostat kanseri, 65 yaşından büyüklerde yaygındır.
Bu çalışmanın amacı, bir seramidaz inhibitörü olan 
(1S,2R)-D-erythro-2-(N-Myristoylamino)-1-phenyl-1-propanol’un
(D-eritro-MAPP) insan prostat kanseri DU-145 hücreleri üzerindeki potansiyel
sitotoksik aktivitesini MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium
Bromide) testi, akış sitometrisi, konfokal ve TEM mikroskopisi kullanarak araştırmaktır.
MTT bulguları D-eritro-MAPP'nin DU-145 hücrelerinde düşük dozlarda toksisiteye
neden olduğunu göstermiştir. Konfokal ve TEM mikroskopi bulguları, hücre
iskeletinde delik oluşumunu, kromatin yoğunlaşmasını, morfolojik değişiklikler
olarak at nalı şeklindeki hücre çekirdeklerini ve hücre zarı üzerindeki
tomurcuklanmaları, çekirdeklerin parçalanmasını, kromatin yoğunlaşmasını ve
ultra yapısal değişiklikler olarak krista kaybını göstermiştir. Akış sitometrisi
bulguları, D-eritro-MAPP'in DU-145 hücrelerine 24 saatlik kısa süreli uygulamasında
apoptozu tetiklediğini göstermiştir. Elde edilen sonuçlara göre, D-eritro-MAPP'in
DU-145 hücrelerinin canlılığını doza bağlı bir şekilde azalttığı söylenebilir.
Bulgularımız, D-eritro-MAPP'in DU-145 hücreleri üzerindeki anti-kanser ve
sitotoksik potansiyelini ortaya koymuştur ve bu ajanın, in vitro ve in vivo
çalışmalardan sonra kanser tedavisi için ilaç geliştirmede kullanılabileceği
belirtilmiştir.

Anahtar Kelimeler

Sfingolipid, DU-145, konfokal mikroskopi

Kaynakça

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