PROSTAT KANSERİ HÜCRE HATTINDA (PC-3) DL-FENİLALANİN VE DL-ALANİN'İN HEDGEHOG YOLU İLE İLİŞKİSİNİN ARAŞTIRILMASI

Abstract

AMAÇ: Bu çalışmada, DL-Fenilalanin ve DL-Alanin’nin PC-3 prostat kanser hücre hattında Hedgehog sinyal yolağı üzerinden etkisinin araştırılması amaçlanmıştır. GEREÇ VE YÖNTEM: DL-Fenilalanin ve DL-Alaninin PC-3 kanser hücrelerinde hücre canlılığına etkisi MTT yöntemi ile belirlemiştir. DL-Fenilalanin için belirlenen IC50 değeri PC-3 ve HEK-293 hücrelerine uygulandı. Total RNA izolasyonu ve cDNA sentezi yapıldı. Hedgehog yolağı ile ilişki anahtar genlerin (SHH, PTCH, SMO, GLI-1) gen ifadesi qPCR (Kantitatif PCR) ile belirlendi. BULGULAR: PC-3 hücreleri üzerinde DL-Alanin sitotoksik etki göstermedi. DL-Fenilalanin hücre canlılığını zamana ve doza bağlı olarak azalttı. DL-Fenilalanin için IC50 değeri 48.saatte 500 μg/mL olarak belirlendi. PC-3 hücrelerinde DL-Fenilalanin uygulamasından sonra tüm genlerin ekspresyonunda aşağı regülasyon, HEK-293 hücrelerinde ise yukarı regülasyon tespit edildi (p<0.05). SONUÇ: DL-Fenilalanin prostat kanseri hücrelerinde Hedgehog yolağını inhibe etmiştir. Bu inhibisyon kontrol grubu HEK-293 hücrelerinde gözlenmemiştir. DL-Fenilalanin prostat kanseri tedavisi için terapötik bir aday olabilir ancak, DL-Alanin’nin böyle bir etkisi bulunmamıştır.

Keywords

• PC-3
• DL-Fenilalanin
• DL-Alanin
• Hedgehog.

INVESTIGATION OF THE RELATIONSHIP OF DL-PHENYLALANINE AND DL-ALANINE WİTH THE HEDGEHOG PATHWAY IN THE PROSTATE CANCER CELL LINE (PC-3)

Abstract

OBJECTIVE: In this study, it was aimed to investigate the effect of DL-Phenylalanine and DL-Alanine on the Hedgehog signaling pathway on the PC-3 prostate cancer cell line. MATERIAL AND METHODS: The effects of DL-Phenylalanine and DL-Alanine on cell viability in PC-3 cancer cells were determined by the MTT method. The IC50 value determined for DL-Phenylalanine was applied to PC-3 and HEK-293 cells. Total RNA isolation and cDNA synthesis were performed. Gene expression of key genes related to the Hedgehog pathway (SHH, PTCH, SMO, GLI-1) was determined by qPCR (Quantitative PCR). RESULTS: DL-Alanine showed no cytotoxic effect on PC-3 cells. DL-Phenylalanine decreased cell viability in a time- and dose-dependent manner. The IC50 value for DL-Phenylalanine was determined to be 500 μg/mL at 48 hours. After DL-Phenylalanine application in PC-3 cells, down-regulation in the expression of all genes was detected, while up-regulation was detected in HEK-293 cells (p<0.05). CONCLUSIONS: DL-Phenylalanine inhibited the Hedgehog pathway in prostate cancer cells. This inhibition was not observed in the control group HEK-293 cells. While DL-Phenylalanine may be a therapeutic candidate for the treatment of prostate cancer, DL-Alanine was not found to have such an effect.

Keywords

• PC-3
• DL-Phenylalanine
• DL-Alanine
• Hedgehog

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