Effects Of Thalidomide And rhGM-CSF on Carbohydrate Metabolism In HL-60 Acute Promyelocytic Leukemia Cell Line

Abstract

The purpose of this study is to investigate the in vitro effect of thalidomide and rhGM-CSF on energy metabolism rate in HL-60 cell lines. HL-60 cells were incubated with radiolabelled glucose for 4 hours. Following incubation, radiolabelled CO2 was isolated and collected in specially designed scintillation vials. The other end-products of carbohydrate catabolism collected via anion-exchange chromatography were analyzed using liquid scintillation. Protein and glycogen levels for each group were also determined. The study results indicated that the use of drugs for 2-days had no significant effect when compared with aerobic and anaerobic controls. However, groups treated with the drugs for 5 days showed significant differences. It was obvious that rhGM-CSF caused HL-60 leukemia cells to use aerobic glycolytic pathway for production of energy. When thalidomide and rhGM-CSF combination therapy results were compared with untreated control cells, it was observed that glycogen consumption was decreased by 50.37%, and CO2 production was increased by 94.03%. When compared with those of anaerobic controls, glycogen consumption and CO2 production rates were found to be decreased by 54.01% and increased by 96.59%, respectively. As a consequence, we found that those cell lines treated with combined use of Thalidomide and rhGM-CSF caused HL-60 leukemia cells to predominantly prefer aerobic glycolytic pathways for energy production.Keywords: Thalidomide, Metabolism, rhGM-CSF, HL-60 cell line ÖZET HL-60 Akut Promyelositik Lösemi Hücre Kültürlerinde Thalidomide ve rhGM-CSF Kullan›lmas›n›n Karbonhidrat Metabolizmas›na EtkileriBu çal›flma, HL-60 hücre kültürlerinde in vitro olarak thalidomide ve rhGM-CSF uygulamas›n›n enerji metabolizmas›na etkilerini araflt›rma amac› ile yap›ld›. HL-60 hücre kültürleri radyoaktif glukozla 4 saat inkübe edildi. Ard›ndan radioaktif iflaretli CO2 özel sintilasyon viallerinde topland›, karbonhidrat katabolizmas›n›n di¤er son ürünleri ise anyon de¤iflim kromotografisi ile ayr›ld› ve s›v› sintilasyon arac›l›¤›yla analiz edildi. Ayr›ca her grubun protein ve glikojen düzeyleri ölçüldü. ‹ki günlük ilaç kullan›m› aerobik ve anaerobik kontrollerde anlaml› bir fark oluflturmazken befl günlük ilaç kullan›m›nda anlaml› fark oldu¤u tespit edildi. rhGM-CSF uygulanan HL-60 lösemi hücre kültürlerinin aerobik glikolitik yola kayd›¤› belirlendi. Thalidomide ve rhGM-CSF kombine tedavisinin sonuçlar›, ilaç verilmeyen aerobik kontrol grubuyla karfl›laflt›r›ld›¤›nda glikojen tüketiminin %50.37 azald›¤›, CO2 üretiminin ise %94.03 oran›nda artt›¤› belirlendi. Kombine ilaç verilen grup anaerobik kontrolle karfl›laflt›r›ld›¤›nda ise, glikojen tüketiminin %54.01 azald›¤›, CO2 üre-timin ise %96.59 artt›¤› belirlendi. Sonuç olarak Thalidomide ve rhGM-CSF kombine kullan›ld›¤›nda, HL-60 lösemi hücrelerinin enerji üretiminde esas olarak aerobik glikolitik yolu tercih ettikleri belirlendi.Anahtar Kelimeler: Talidomit, Metabolizma, rhGM-CSF, HL-60 hücre serisi ULUSLARARASı HEMATOLOJI-ONKOLOJI DERGISI ARTICLE