Glutaraldehit Kaplı Manyetik Nanopartiküle İdarubisin Tutturulması ve HL-60 Hücre Hattında Etkisinin İncelenmesi

Abstract

İdarubisin, meme kanseri ve akut lösemi tedavisinde sıklıkla kullanılan kemoterapötik bir ilaçtır. Bu çalışmada, yüksek stabilite ve düşük toksisiteye sahip bir ilaç hazırlamak için idarubisini glutaraldehit (GA) kaplı manyetik nanopartiküllere (MNP-GA) immobilize edilmesi amaçlanmıştır. Manyetik nanopartiküller kolay sentez edilmesi, düşük maliyetli olması ve toksik olmamasından dolayı tercih edilmiştir. Çalışmada magnetit (Fe3O4) nanoparçacıkları hazırlanmış, glutaraldehit ile kaplanmış, Fourier transform kızılötesi spektroskopisi, X-ışını kırınım deseni ve Konvansiyonel transmisyon elektron mikroskobu yöntemleri ile karakterizasyon işlemleri yapılmış ve idarubisin bağlanmıştır. İdarubisine bağlı MNP-GA ve serbest idarubisinin HL-60 hücre hatları üzerindeki sitotoksik etkileri MTT ve ATP testleri ile belirlendi ve IC50 değerleri hesaplandı. Apoptoz durumu flow sitometrisi ile değerlendirildi ve MDR1, Puma, NOXA, BAX, Survivin ve BCL-2 genlerinin ekspresyonu polimeraz zincir reaksiyonu (PCR) yöntemi ile ölçüldü. MNP kullanımı ile IC50'nin 5 ile 7 kat arasında azaldığı tespit edilmiştir. PCR testlerinde apoptotik genlerin ekspresyonları artarken, MNP kullanımında MDR1 ve anti-apoptotik genlerin ekspresyonları azaldı. Akım sitometri ölçümlerinde apoptoz artışı saptandı. MNP sistemlerinin kullanımı, ilacın kontrollü salınımını sağladığı ve yapısı gereği hücreden çıkışını engellediği için ilaç direncini azaltmıştır.

Keywords

• Akut Promyelositik Lösemi
• Glutaraldehit
• HL-60 Hücre Hattı
• İdarubisin
• Manyetik Nanopartiküller

Attachment of Idarubicin to Glutaraldehyde-coated Magnetic Nanoparticle and Investigation of its Effect in HL-60 Cell Line

Abstract

Idarubicin is a chemotherapeutic drug frequently used to treat breast cancer and acute leukemia. This study aimed to immobilize idarubicin on glutaraldehyde (GA)-coated magnetic nanoparticles (MNP-GA) to prepare a drug with high stability and low toxicity. We prefreed MNPS because of their easy synthesis, low cost, and non-toxicity. In the study, magnetite (Fe3O4) nanoparticles were prepared, coated with glutaraldehyde, characterization processes were performed with Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction pattern (XRD), and Conventional transmission electron microscopy (C-TEM) methods, and idarubicin (IDA) was bound. The cytotoxic effects of idarubicin-bound MNP-GA and free idarubicin on HL-60 cell lines were determined by MTT and ATP tests, and IC50 values were calculated. Flow cytometry was used to evaluate apoptosis status, and the expression of MDR1, Puma, NOXA, BAX, Survivin, and BCL-2 genes were measured by the polymerase chain reaction (PCR). It was found that the IC50 decreased between 5 and 7 times with the use of MNP. In PCR tests, the expressions of apoptotic genes increased, while the expressions of MDR1 and anti-apoptotic genes were decreased in the use of MNP. Apoptosis was found to be increased in flow cytometry measurements. The use of MNP systems has reduced drug resistance since it provides controlled release of the drug and prevents its exit from the cell due to its structure.

Keywords

• Acute Promyelocytic Leukemia
• Glutaraldehyde
• HL-60 Cell Line
• Idarubicin
• Magnetic Nanoparticles

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