Aprepitantın İnsan Glioblastoma U87MG Hücreleri üzerinde Antiproliferatif ve Apoptotik Etkileri

Glioblastoma multiforme, gliomalar arasında en malign özellikte olup, görülme sıklığı oldukça yüksektir. Hızlı çoğalma, beyin dokusuna invazyon ve migrasyon gibi agresif özelliklere sahiptir. Bu nedenle son yıllarda glioblastoma kemoterapisinde, yeni tedavi hedeflerinin gelişmesini üzerine çalışmalar yoğunlaşmıştır. Substans P, nörokinin-1 (SP/nk-1) reseptör sistemi, kanser gelişimi ve kanser metastazında önemli bir rol oynamaktadır. Bu nedenle nk-1 reseptörleri, glioblastoma tedavisinde ve nk-1 reseptör antagonistlerinin gelişiminde yeni tedavi hedefleri arasında yer almaktadır. Bu çalışmada, klinikte özellikle kemoterapi sırasında antiemetik amaçla kullanılan, bir nk-1 reseptör antagonisti olan aprepitantın, insan malignant glioblastoma U87MG hücre hattında antiproliferatif ve apoptotik etkileri araştırılmıştır. Çalışmada MTT yöntemi ve gerçek zamanlı hücre analiz sistemi (RTCA DP) ile hücre proliferasyon analizleri, anneksin V-PI ve kaspaz 3 flow sitometri analizleri ile de apoptotik etkiler çalışılmıştır. MTT analizine göre aprepitantın konsantrasyon ve zamana artışına bağlı olarak U87MG hücreleri üzerinde sitotoksik etki yaptığı ve U87MG hücre proliferasyonunu azalttığı görülmüştür. Gerçek zamanlı analiz sisteminden elde edilen hücre indeks değerlerine göre de, 50 µM ve üzeri aprepitant konsantrasyonlarında hücre proliferasyonunun azaldığı ve sırasıyla IC50 değerlerinin 24. saatte 76.9 µM, 48. saatte 62.5 µM ve 72. saatte de 59.2 µM olarak belirlenmiştir. Flow sitometri analiz sonuçlarına göre aprepitantın 24. saatteki erken ve geç apoptotik etkisi konsantrasyon artışına bağlı olarak artış göstermiştir. Özellikle IC25 ve IC50 aprepitant değerlerinde aktif kaspaz-3 değeri kontrole göre, az da olsa artmıştır. Sonuç olarak aprepitantın malignant glioblastoma U87MG hücrelerinde önemli antiproliferatif ve apoptotik etkileri belirlenmiş olup, nk-1 reseptör antagonistlerinin yeni bir terapötik hedef olabileceği düşünülmektedir.

Antiproliferative and Apoptotic effects of Aprepitant on Human Glioblastoma U87MG Cells

Glioblastoma multiforme, is the most malignant glioma among other gliomas with high incidence. It has aggresive properties such as high proliferation, invasion and migration rates. Therefore, recently studies have focused on the development of new therapeutic targets in glioblastoma chemotherapy. Substance P, neurokinin-1 (SP/nk-1) receptor system has an important role in cancer development and metastasis. Therefore, nk-1 receptors are new therapeutic targets for glioblastoma therapy and development of nk-1 receptor antagonists. In this study, antiproliferative and apoptotic effects of aprepitant which is an antiemetic drug commonly using in chemotherapy on human U87MG glioblastoma cells were investigated. Antiproliferative effects of aprepitant were evaluated by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and Real Time Analyses System (RTCA DP). Also apoptotic effects of aprepitant were evaluated by annexin-VPI and caspase-3 activity by using flow cytometer. According to the cell index values obtained from real-time cell analysis system, with 50 µM and higher concentrations, cell proliferation was decreased and IC50 values were determined as 76.9, 62.5 and 59.2 µM for 24, 48 and 72 hours respectively. Aprepitant showed significant antiproliferative effects on U87MG cells. Depending on increasing aprepitant concentrations, both early and late apoptotic cell ratios were increased. In addition, caspase-3 activation was slightly increased with IC25 and IC50 concentrations of aprepitant according to the control group. As a result, aprepitant showed important antiproliferative and apoptotic effects on U87MG cells. In conclusion, nk-1 receptor antagonists may have a potential therapeutic role for human glioblastoma chemotherapy.

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