Elektroporasyon İşlemi Sırasında Oluşan Hücre Ölümünde Darbe Frekansının Önemi

Abstract

Tersinir elektroporasyon (EP), darbeli elektrik alanlarının uygulanmasıyla hücre zarının geçirgenliğinin arttırıldığı biyofiziksel bir yöntemdir. Kanser tedavisinde, EP tümör hücrelerinde antikanser ilaçlarının alımını arttırmak, dolayısıyla sitotoksik etkilerini yükseltmek için kullanılır, ancak hücre ölümüne de yol açabilmektedir. Bu çalışmanın amacı tersinir EP uygulamasında meydana gelen hücre ölümünde darbe frekansının etkisini incelemektir. Çalışmada HepG2 karaciğer kanser hücreleri kullanıldı. Hücreler 200 Hz, 1 kHz, 5 kHz, 10 kHz ve 20 kHz frekanslarında 70-1250 V/cm elektrik alana sahip, 100μs süreli 8 kare dalga elektrik darbesine maruz bırakıldı. Elektrik maruziyetinden sonra hücre canlılığı MTT tahlili ile belirlendi. Çalışmamızın MTT bulgularına göre, EP’de uygulanan darbe frekansı arttıkça HepG2 kanser hücrelerinde inhibisyon azalmaktadır. EP uygulanan HepG2 hücrelerinde en yüksek inhibisyon, 1000 V/cm elektrik alanında ve 200 Hz'lik frekansta meydana geldi. 10 kHz ve 20 kHz frekanslarında uygulanan düşük elektrik alanlar (70 V/cm, 250 V/cm) hücre canlılığında azalmaya neden olmadı. Düşük elektrik alanlarında 1 kHz frekansına göre 5 kHz frekansından daha fazla hücre ölümü gerçekleşse de, yüksek elektrik alanlarında 1 kHz frekansında daha yüksek oranda hücre ölümü tespit edildi. Sonuçlarımız, elektroporasyonda düşük frekanslardaki darbelerin daha fazla hücre inhibisyonuna neden olabildiğini, bu yüzden yüksek frekanslı darbelerin kullanımı daha uygun olabileceğini göstermektedir.

Keywords

• Darbe Frekans
• Elektrokemoterapi
• Elektroporasyon
• HepG2 Hücreleri
• Hücre İnhibisyonu

The Importance Of Pulse Frequency in Cell Death During Electroporation Process

Abstract

Reversible electroporation (EP) is a biophysical method in which the permeability of the cell membrane is enhanced by the application of pulsed electric fields. In cancer treatment, EP is used to increase the uptake of anticancer drugs in tumor cells, thus increasing their cytotoxic effects, but it can also lead to cell death. The aim of this study is to examine the effect of pulse frequency on cell death in reversible EP applications. HepG2 liver cancer cells were used in the study. The cells were exposed to 8 square wave electrical pulses of 100 μs duration with an electric field of 70-1250 V/cm at frequencies of 200 Hz, 1kHz, 5kHz, 10kHz and 20kHz. Cell viability after electrical exposure was determined by MTT assay. According to the MTT findings of our study, inhibition in HepG2 cancer cells decreases as the pulse frequency applied in EP increases. The highest inhibition in EP applied HepG2 cells occurred at 1000 V/cm electric field and 200 Hz frequency. Low electric fields (70 V/cm, 250 V/cm) applied at 10 kHz and 20kHz frequencies did not cause a decrease in cell viability. Although more cell death occurred at a frequency of 5 kHz than at a frequency of 1kHz in low electric fields, a higher rate of cell death was detected at a frequency of 1kHz in high electric fields. Our results show that low-frequency pulses in electroporation can cause more cell inhibition, so the use of high-frequency pulses may be more appropriate.

Keywords

• Cell Inhibition
• Electrochemotherapy
• Electroporation
• HepG2 Cells
• Pulse Frequency

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