Özet
Dayanak
Üçlü negatif meme kanseri (TNBC), kötü prognoz ve sınırlı tedavi seçenekleri ile ilişkili agresif bir kanser türüdür. Hedeflenebilir reseptörlerin bulunmaması nedeniyle, geleneksel kemoterapi birincil tedavi yaklaşımı olmaya devam etmektedir. ERCC1, karboplatin gibi platin bazlı ajanlar tarafından hasar gören DNA’yı onarmaktan sorumlu olan nükleotid eksizyon onarım sisteminin kritik bir bileşenidir. Isı şoku yanıtı (HSR), kanser hücrelerinin hayatta kalmasını destekleyen, stres ile indüklenen temel bir savunma mekanizmasıdır.
Amaçlar
Bu çalışma, KNK437 aracılı HSR inhibisyonunun TNBC hücre hattı MDA-MB-231’de ERCC1 gen ekspresyonu ve karboplatin duyarlılığı üzerindeki etkilerini araştırmayı amaçlamaktadır.
Yöntemler
Karboplatin ve KNK437’nin IC50 değerleri WST-8 sitotoksisite testiyle belirlenmiştir. ERCC1 gen ekspresyon düzeyleri gerçek zamanlı kantitatif polimeraz zincir reaksiyonu ile ölçülmüştür. Karboplatin ve KNK437 tarafından indüklenen apoptotik ve nekrotik hücre ölümü, FITC-Annexin V testi kullanılarak akış sitometrisi ile saptanmıştır.
Bulgular
Karboplatin ve KNK437’nin IC50 değerleri sırasıyla 247,5 µM ve 89,74 µM olarak hesaplanmıştır. Karboplatin veya KNK437 monoterapisi ERCC1 ekspresyonunu sırasıyla %42,8 ve %49,5 oranında azaltırken, bunların kombine uygulaması %54,9 oranında bir azalmaya neden olmuştur. Ayıca, kombine uygulama karboplatinin tek başına kullanımına kıyasla toplam hücre ölümünü %34,1 oranında belirgin şekilde artırmıştır. İlginç olarak, karboplatin tarafından indüklenen nekroz, KNK437 ile kombine uygulamada apoptoza doğru bir kayma göstermiştir, ve bu durum ışık mikroskobu ve akış sitometrisi ile doğrulanmıştır.
Sonuç
Bulgularımız, KNK437 aracılı HSR inhibisyonunun TNBC hücrelerinde karboplatin duyarlılığını artırdığını ve ERCC1 gen ekspresyonunu aşağı regüle ettiğini göstermektedir. TNBC’nin agresif doğası ve sınırlı tedavi seçenekleri göz önüne alındığında, sonuçlarımız KNK437’nin karboplatin ile kombine edildiğinde, özellikle karboplatin kaynaklı nekrozun inflamasyonla ilişkili komplikasyonlara katkıda bulunduğu durumlarda, terapötik avantajlar sunabileceğini göstermektedir.
Anahtar Kelimeler:
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