Simge ÜNAY, Mehmet Dinçer BİLGİN

Ht-29 Hücrelerinde 5-ALA Kullanılarak Tümör Floresan Görüntüleme Yöntemi

Amaç: 5-aminolevulinik asit olarak da bilinen 5-ALA, hemoglobinin önemli bir bileşeni olan heme ve vücuttaki çeşitli enzimlerin biyosentezinde çok önemli bir rol oynayan doğal olarak oluşan bir amino asittir. Mitokondride glisin ve süksinil-CoA'dan sentezlenir. Spesifik olarak, fotodinamik tanı (PDD) ve fotodinamik terapi (PDT) adı verilen teknikte bir ışığa duyarlılaştırıcı olarak kullanılır. Fotodinamik tanıda (PDD), 5-ALA tümör hücreleri veya kanser öncesi lezyonlar gibi belirli doku veya hücrelerde seçici olarak birikir. Spesifik bir dalga boyundaki ışığa maruz kaldığında, biriken 5-ALA, bu hedef alanlarda floresana neden olarak beyin cerrahisi veya üroloji gibi cerrahi prosedürler sırasında gelişmiş görselleştirme ve algılamaya olanak tanır. Bu çalışmanın amacı, kolon kanserinde 5-ALA kullanılarak optimal fotodinamik tanı koşullarını belirlemektir. Yöntem: HT-29 hücre hattına 3 saat farklı 5-ALA (100, 200, 300, 500, 1000, 1500 μM) konsantrasyonları uygulanmıştır. Uygulanan hücrelerde hücre canlılığı, floresans şiddeti, apoptoz analizleri yapıldı. Bulgular: Kontrol grubu ile düşük 5-ALA dozları (100,200 ve 300 μM) arasında hücre canlılığı açısından fark yokken, yüksek 5-ALA dozlarında (1000 ve 1500 μM) anlamlı fark bulundu (p<,0001) 5-ALA dozları floresan şiddeti ile paralel olarak artmış ve en yüksek floresans şiddeti 1500 μM 5-ALA'da olmuştur (p<,0001). Apoptoz/ölü oranı, floresans yoğunluğunun en yüksek olduğu 1000 μM ve 1500 μM 5-ALA'da anlamlı olarak en yüksek olduğu gösterildi (p<0.05). Sonuç: HT-29 hücrelerinde 5-ALA konsantrasyonunun optimum dozu 500 μM olduğu belirlendi. 5-ALA'nın yüksek konsantrasyonlarının HT29 hücrelerinde apoptoza neden olduğu gösterilmiştir.

Anahtar Kelimeler:

5-aminolevulinik asit (ALA), Floresans görüntüleme, Tümör hücreleri, Apoptoz, Fotodinamik Tanı

Tumor Florescence Imaging Method Using 5-ALA in Ht-29 Cells

Objective: 5-ALA, also known as 5-aminolevulinic acid, is a naturally occurring amino acid that plays a crucial role in the biosynthesis of heme, a vital component of hemoglobin and various enzymes in the body. Specifically, it is used in technique called photodynamic diagnosis (PDD) and photodynamic therapy (PDT) as a photosensitizer. When exposed to a specific wavelength of light, the accumulated 5-ALA causes fluorescence in these target areas, allowing for enhanced visualization and detection during surgical procedures, such as in neurosurgery or urology. The purpose of this study was to evaluate the conditions for optimal photodynamic diagnosis using 5-ALA in colon cancer. Methods: HT-29 cell line which was administered different 5-ALA (100, 200, 300, 500, 1000, 1500 μM) concentrations for 3 hours incubation time, were performed on cell viability, fluorescence intensity, apoptosis analysis. Results: While there was no difference in cell viability between the control group and low 5-ALA doses (100,200 and 300 μM), a significant difference was found at higher 5-ALA doses (1000 and 1500 μM) (p<,0001). 5-ALA doses increased in parallel with the fluorescence intensity, and the highest fluorescence intensity was at 1500 μM 5-ALA (p<,0001). The apoptosis/dead ratio was significantly showed to be highest at 1000 μM and 1500 μM 5-ALA which had the highest fluorescence intensity (p<0.05). Conclusion: the optimum dose of 5-ALA concentration was determined to be 500 μM in HT-29 cells. High concentrations of 5-ALA have been shown to cause apoptosis in HT29 cells.

Keywords:

5-aminolevulinic acid (ALA), Florescence Imaging, Tumor cells, Apoptosis, Photodynamic Diagnosis,

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