Investigation of 5-Fluorouracil-Induced Cell Death in HCT116 and HT29 Colon Cancer Cells by Fourier Transform Infrared Spectroscopy

Abstract

Colon cancer is a common and highly lethal type of cancer all over the world. As with all cancers, monitoring the course of colon cancer is critical. 5-Fluorouracil (5-FU) is an anticancer drug frequently used in colon cancers and causes some biochemical and molecular differences in cells during cell death. In this study, after incubation of HCT116 and HT29 colon cancer cells with 5-FU chemotherapy drug, Fourier transform infrared (FTIR) spectroscopy was used to detect the biochemical change caused by 5FU on cells. The concentration (IC50) of 5-FU at which percent inhibition equals 50 was determined using the MTT method. FTIR measurements were taken after the IC50 value was determined. Accordingly, the IC50 value of 5-FU was found to be 12.69 µg/ml for HCT116 cells and 10.10 µg/ml for HT29 cells, respectively. FTIR spectroscopy showed changes in IR biomarkers such as the increase in lipid content, which is said to be indicators of cell death caused by 5-FU, and the nucleic acid ratio, the α-helix and β-sheet secondary structures of total cellular proteins, and the reduction of amino acid residues, and changes in collagen conformation. As a result, examining cancer cell death with FTIR, which is both a cheap and fast method, can be used as an alternative method.

Keywords

• Colon cancer
• 5-fluorouracil

HCT116 ve HT29 Kolon Kanseri Hücrelerinde 5-Florourasil Kaynaklı Hücre Ölümünün Fourier Dönüşümlü Kızılötesi Spektroskopisi ile İncelenmesi

Abstract

Kolon kanseri, tüm dünyada yaygın olan ve yüksek oranda ölümcül bir kanser tipidir. Tüm kanserlerde olduğu gibi kolon kanserinin seyrinin izlenmesi kritik bir öneme sahiptir. 5-Florourasil (5-FU) kolon kanserlerinde sıklıkla kullanılan bir antikanser ilaçtır ve hücre ölümü esnasında hücrelerde bir takım biyokimyasal ve moleküler farklılıklara neden olmaktadır. Bu çalışmada HCT116 ve HT29 kolon kanseri hücreleri 5-FU kemoterapi ilacı ile inkübe edildikten sonra, 5FU’nun hücreler üzerinde oluşturduğu biyokimyasal değişikliğin tespiti için Fourier transform kızılötesi (FTIR) spektroskopisi kullanılmıştır. 5-FU'nun inhibisyon yüzdesinin 50'ye eşit olduğu konsantrasyonu (IC50), MTT yöntemi kullanılarak belirlendi. IC50 değeri saptandıktan sonra FTIR ölçümleri alındı. Buna göre, 5-FU IC50 değeri sırasıyla HCT116 hücreleri için 12,69 µg/ml, HT29 hücreleri için 10,10 µg/ml bulunmuştur. 5-FU’nun sebep olduğu hücre ölümünün göstergeleri olduğu söylenen lipit içeriğindeki artış ile nükleik asit oranı, total hücresel proteinlerin α-sarmal ve β-yaprak ikincil yapıları ve aminoasit kalıntılarının azalması, kollajen konformasyonundaki değişiklikler gibi IR biyobelirteçlerindeki değişiklikler FTIR spektroskopisi ile gösterilmiştir. Sonuç olarak hem ucuz ve hem de hızlı bir yöntem olan FTIR ile kanser hücre ölümünün incelenmesi alternatif bir yöntem olarak kullanılabilir.

Keywords

• Kolon kanseri
• 5-florourasil
• FTIR
• Colon cancer
• 5-fluorouracil

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