Erythroid Differentiation Inducer, Hemin Inhibits Cyclic AMP Production in K562 Cells

Year 2011, Volume: 3 Issue: 1, 54 - 58, 01.04.2011

Bahire Küçükkaya

Abstract

Objective: This study investigated the intracellular cyclic AMP concentrations in hemin-induced and uninduced K562 cell line. Methods: K562 cell line were grown in RPMI medium supplemented with 10% fetal calf serum FCS , 100 IU/ml penicillin, 100 μg/ml streptomycin, 25 μg/ml amphotericin B and 2 mM L-glutamin at 37 oC in humidified air containing 5% CO2. A Trypan blue stain viability test was performed for produced K562 cells, and those used to obtain cell pellets from 1st day to 6th day, under the the following conditions: without hemin treatment as a control group and with hemin treatment as an experimental group. All data were statistically analyzed using one-way ANOVA, followed by Student`s t test. Results: Treatment of K562 cells with hemin leads to inhibition of cell proliferation. Cyclic AMP levels of K562 cells that were treated with hemin are decreased in a time dependent manner, although the hemin-induced cells proliferated. Intracellular cyclic AMP levels of hemininduced K562 cells decreased, while the control cells had stable cyclic AMP concentrations. Conclusion: On the basis of these results, it is recommended that further data collection is needed to analyze the mechanism by which intracellular cyclic AMP levels are regulated in K562 cells treated with hemin or other erythroid inducers.

Keywords

cAMP, hemin, K562, erythroid differentiation, cellular signaling

K562 Hücrelerinde Eritroid Farklılaşmayı Uyaran Hemin, Siklik AMP Oluşumunu Baskılar

Abstract

Amaç: Bu çalışmada heminle indüklenen ve indüklenmeyen K562 hücrelerinde, hücre içi siklik AMP düzeyleri incelendi. Yöntemler: K562 hücreleri %10 fetal dana serumu, 100 IU/ml penisilin, 100 μg/ml streptomisin, 25 μg/ml amfoterisin B ve 2 mM L-glutamin içeren RPMI-1640 içerisinde ve nemli % 5'lik karbondioksit etüvünde, 37 oC'da çoğaltıldı. Çoğaltılan bu hücrelerde Tripan mavisi boyama canlılık testi yapıldı. Hemin ile muamele edilen deney grubu ve hemin ile muamele edilmeyen kontrol grubu hücrelerden birinci günden itibaren altıncı güne kadar hücre peletleri elde edildi. Elde edilen bu hücre peletlerinde siklik AMP konsantrasyonları, siklik AMP Enzim ‹mmuno Analiz sistemi kullanılarak ölçüldü. Tüm veriler Student's t-testi' ni takiben tek-yol ANOVA ile istatistiksel olarak analiz edildi. Bulgular: K562 hücrelerinin heminle muamelesi, hücre çoğalmasının baskılanmasına neden olmuştur. Heminle muamele edilen K562 hücrelerinin çoğalmasına rağmen siklik AMP düzeyleri zamana bağlı olarak azalmıştır. Heminle indüklenen K562 hücrelerinin hücre içi siklik AMP düzeyleri azalırken, kontrol hücrelerinin siklik AMP düzeyleri kararlı kalmıştır. Sonuç: Bu sonuçlara dayanarak, diğer eritroid indükleyiciler veya hemin ile muamele edilen K562 hücrelerinde hücre içi siklik AMP düzeylerinin hangi mekanizmayla düzenlendiğ ini anlamak için daha çok bilgiye gerek vardır.

Keywords

cAMP, hemin, K562, eritroid farklılaşma, hücresel sinyal iletimi

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