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

Yıl 2010, Cilt: 2 Sayı: 2, 32 - 36, 01.08.2010

Öz

Objective: This study investigated the intracellular cyclic AMP concentrations in hemin-induced and uninduced K562 cell line. Method: K562 cell line were grown in RPMI medium supplemented with 10% fetal calf serum FCS , 100 IU/ml penicillin, 100 Ìg/ml streptomycin, 25 Ìg/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 1 day to 6 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 are 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.

Kaynakça

  • )Pomerance M, Abdullah HB, Kamerji S, Correze C, Blondeau JP.: Thyroid- stimulating hormone and cyclic AMP activate p38 mitogen-activated protein kinase cascade. Involvement of protein kinase A, rac1, and reactive oxygen species. J Biol Chem. 2000; 275:40539-40546.
  • )Cook SJ, McCormick F.: Inhibition by cAMP of Ras Dependent Activation of Raf. Science. 1993; 262: 1069-1072. Takanaga H, Yoshitake T, Hara S, Yamasaki C, Kunimoto M.: cAMP-induced astrocytic differentia- tion of C6 glioma cells is mediated by autocrine inter- leukins-6. J Biol Chem. 2004; 279:15441-15447.
  • )Miyata A, Arimura.A, Dahl RR, Minamino NN, Uehara A, Jiang L, Culler MD, Coy DH.: Isolation of a novel 38 residue-hypothalamic polypeptide which stimulates adenylate cyclase in pituitary cells. Biochem Biophys Res Commun. 1989;164:567-574.
  • )Basille M, Vaudry D, Coulouran Y, Jegou S, Lihramann I, Fournier A, Vaudry H, Gonzalez B.: Comparative distribution of Pituitary adenylate cyclase-activating polypeptide (PACAP) binding sites and PACAP receptor mRNAs in the rat brain during development. J Comp Neurol. 2000;425:495-509.
  • )Jaworski DM, Proctor MD.: Developmental regulation of pituitary adenylate cyclase-activating polypeptide and PAC(1) receptor mRNA expression in the rat cen- tral nervous system. Dev Brain Res. 2000;120:27-39.
  • )Vallejo I, Vallejo M.: Pituitary adenylate cyclase activating polypeptide induces astrocyte differentiation of precursor cells from developing cerebral cortex. Mol Cell Neurosci. 2002;21:671-683.
  • )Grdisa M, White MK.: Erythrocytic differentiation and glyceraldehydes - 3 - phosphate dehydrogenase expression are regulated by protein phosphorylation and cAMP in HD3 cells. Int J Biochem Cell Biol. 2000;32:589-595.
  • )Andersson LC, Jokinen M, Gahmberg CG.: Induction of erythroid differentiation in the human leukemia cell line K562. Nature. 1979;278:364-365.
  • )Küçükkaya B, Öztürk L, Yalçıntepe L.: Nitric oxide levels during erythroid differentiation in K562 cell line. Indian J Biochem Biophys. 2006;43:251-253.
  • )Cioe L, McNab A, Hubbell HR, Meo P, Curtis P, Rovera G.: Differential expression of the globin genes in human leukemia K562(S) cells induced to differentiate by hemin or butyric acid. Cancer Res. 1981;41:237-243.
  • )Pardoe IU, Grewal KK, Baldeh MP, Hamid J, Burness AT.: Persistent infection of K562 cells by encephalomy- ocarditis virus. J Virol. 1990; 64:6040-6044.
  • )Ikuta T, Ausenda S, Cappellini MD.: Mechanism for fetal globin gene expression role of the soluble guanylate cyclase-cyclic GMP-dependent protein kinase pathway. Proc Natl Acad Sci U S A. 2001; 98:1847-1852.
  • )Francis SH, Corbin JD.: Structure and function of cyclic-nucleotide dependent protein kinases. Annu Rev Physiol. 1994;56:237-272.
  • )Ghil S, Choi JM, Kim SS, Lee YD, Liao Y, Birnbaumer L, Suh-Kim H.: Compartmentalization of protein kinase A signaling by the heterotrimeric G protein Go. Proc Natl Acad Sci U S A. 2006;103:19158-19163.
  • )Lozzio CB, Lozzio BB.: Human chronic myelogenous leukemia cell line with positive Philadelphia chromosome. Blood. 1975;45:321-334.
  • )Rowley PT, Ohlsson-Wilhelm BM, Farley BA, LaBella S.: Inducers of erythroid differentiation in K562 human leukemia cells. Exp Hematol. 1981;9:32-37.
  • )Kitanaka A, Waki M, Kamano H, et al.: Antisense src expression inhibits proliferation and erythropoietin- induced erythroid differentiation of K562 human leukemia cells. Biochem Biophys Res. Commun. 1994;201:1534-1540.
  • )Rutherford TR, Clegg JB, Weatherall DJ.: K562 human leukaemic cells synthesise embryonic haemoglobin in response to haemin. Nature. 1979;280:164-165. Morgan L, Jessen KR, Mirsky R.: The effects of camp on differentiation of cultured Schwann cells: Progression from an early phenotype (04+) to a myelin phenotype (Po+, GFAP- N-CAM-, NGF- Receptor-) Depends on Growth inhibition. The J Cell Biol. 1991;112:457-467.
  • )Dumont JE, Jauniaux JC, Roger PP.: The cyclic AMP mediated stimulation of cell proliferation. Trends Biochem Sci. 1989 ;14:67-71.
  • )Chen J, Iyengar R.: Suppression of Ras-induced transformation of NIH 3T3 cells by activated G·s. Science. 1994;263:1278-1281.
  • )Vallar L.: Oncogenic role of heterotrimeric G proteins. Cancer Surv. 1996;27:325-338. Nahorski SR.: Pharmacology of intracellular signalling pathways. Br J Pharmacol. 2006; 147:S38-S45.
  • )Sgambati SA, Zarif A, Basson MD.: Octreotide differentially modulates human Caco-2 intestinal epithelial cell proliferation and differentiation by decreasing intracellular cAMP. Regul Pept. 1996; 61: 219-227.
  • )Inoue A, Kuroyanagi Y, Terui K, Moi P, Ikuta T.: Negative regulation of globin gene expression by cyclic AMP-dependent pathway in erythroid cells. Exp Hematol. 2004;32:244-253.
  • )Cokic VP, Andric SA, Stojilkovic SS, Noguchi CT, Schechter AN.: Hydroxyurea nitrosylates and activates soluble guanylyl cyclase in human erythroid cells. Blood. 2008; 111:1117-1123. Küçükkaya B,.Arslan DO, Kan B.: Role of G proteins and ERK activation in hemin-induced erythroid differentiation of K562 cells. Life Sci. 2006;78:1217- 1224.

K562 Hücrelerinde Eritroid Farklılaşmayı Uyaran Hemin, Siklik AMP Oluşumunu Baskılar

Yıl 2010, Cilt: 2 Sayı: 2, 32 - 36, 01.08.2010

Öz

Amaç: Bu çalışmada heminle indüklenen ve indüklenmeyen K562 hücrelerinde siklik AMP düzeylerinin incelenmesi planlandı. Gereç ve Yöntemler: K562 hücreleri % 10 fetal dana serumu, 100 IU/ml penisilin, 100 Mg/ml streptomisin, 25 Mg/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 siklik AMP düzeylerinin hangi mekanizmayla düzenlendiğini anlamak için daha çok bilgiye gerek vardır.

Kaynakça

  • )Pomerance M, Abdullah HB, Kamerji S, Correze C, Blondeau JP.: Thyroid- stimulating hormone and cyclic AMP activate p38 mitogen-activated protein kinase cascade. Involvement of protein kinase A, rac1, and reactive oxygen species. J Biol Chem. 2000; 275:40539-40546.
  • )Cook SJ, McCormick F.: Inhibition by cAMP of Ras Dependent Activation of Raf. Science. 1993; 262: 1069-1072. Takanaga H, Yoshitake T, Hara S, Yamasaki C, Kunimoto M.: cAMP-induced astrocytic differentia- tion of C6 glioma cells is mediated by autocrine inter- leukins-6. J Biol Chem. 2004; 279:15441-15447.
  • )Miyata A, Arimura.A, Dahl RR, Minamino NN, Uehara A, Jiang L, Culler MD, Coy DH.: Isolation of a novel 38 residue-hypothalamic polypeptide which stimulates adenylate cyclase in pituitary cells. Biochem Biophys Res Commun. 1989;164:567-574.
  • )Basille M, Vaudry D, Coulouran Y, Jegou S, Lihramann I, Fournier A, Vaudry H, Gonzalez B.: Comparative distribution of Pituitary adenylate cyclase-activating polypeptide (PACAP) binding sites and PACAP receptor mRNAs in the rat brain during development. J Comp Neurol. 2000;425:495-509.
  • )Jaworski DM, Proctor MD.: Developmental regulation of pituitary adenylate cyclase-activating polypeptide and PAC(1) receptor mRNA expression in the rat cen- tral nervous system. Dev Brain Res. 2000;120:27-39.
  • )Vallejo I, Vallejo M.: Pituitary adenylate cyclase activating polypeptide induces astrocyte differentiation of precursor cells from developing cerebral cortex. Mol Cell Neurosci. 2002;21:671-683.
  • )Grdisa M, White MK.: Erythrocytic differentiation and glyceraldehydes - 3 - phosphate dehydrogenase expression are regulated by protein phosphorylation and cAMP in HD3 cells. Int J Biochem Cell Biol. 2000;32:589-595.
  • )Andersson LC, Jokinen M, Gahmberg CG.: Induction of erythroid differentiation in the human leukemia cell line K562. Nature. 1979;278:364-365.
  • )Küçükkaya B, Öztürk L, Yalçıntepe L.: Nitric oxide levels during erythroid differentiation in K562 cell line. Indian J Biochem Biophys. 2006;43:251-253.
  • )Cioe L, McNab A, Hubbell HR, Meo P, Curtis P, Rovera G.: Differential expression of the globin genes in human leukemia K562(S) cells induced to differentiate by hemin or butyric acid. Cancer Res. 1981;41:237-243.
  • )Pardoe IU, Grewal KK, Baldeh MP, Hamid J, Burness AT.: Persistent infection of K562 cells by encephalomy- ocarditis virus. J Virol. 1990; 64:6040-6044.
  • )Ikuta T, Ausenda S, Cappellini MD.: Mechanism for fetal globin gene expression role of the soluble guanylate cyclase-cyclic GMP-dependent protein kinase pathway. Proc Natl Acad Sci U S A. 2001; 98:1847-1852.
  • )Francis SH, Corbin JD.: Structure and function of cyclic-nucleotide dependent protein kinases. Annu Rev Physiol. 1994;56:237-272.
  • )Ghil S, Choi JM, Kim SS, Lee YD, Liao Y, Birnbaumer L, Suh-Kim H.: Compartmentalization of protein kinase A signaling by the heterotrimeric G protein Go. Proc Natl Acad Sci U S A. 2006;103:19158-19163.
  • )Lozzio CB, Lozzio BB.: Human chronic myelogenous leukemia cell line with positive Philadelphia chromosome. Blood. 1975;45:321-334.
  • )Rowley PT, Ohlsson-Wilhelm BM, Farley BA, LaBella S.: Inducers of erythroid differentiation in K562 human leukemia cells. Exp Hematol. 1981;9:32-37.
  • )Kitanaka A, Waki M, Kamano H, et al.: Antisense src expression inhibits proliferation and erythropoietin- induced erythroid differentiation of K562 human leukemia cells. Biochem Biophys Res. Commun. 1994;201:1534-1540.
  • )Rutherford TR, Clegg JB, Weatherall DJ.: K562 human leukaemic cells synthesise embryonic haemoglobin in response to haemin. Nature. 1979;280:164-165. Morgan L, Jessen KR, Mirsky R.: The effects of camp on differentiation of cultured Schwann cells: Progression from an early phenotype (04+) to a myelin phenotype (Po+, GFAP- N-CAM-, NGF- Receptor-) Depends on Growth inhibition. The J Cell Biol. 1991;112:457-467.
  • )Dumont JE, Jauniaux JC, Roger PP.: The cyclic AMP mediated stimulation of cell proliferation. Trends Biochem Sci. 1989 ;14:67-71.
  • )Chen J, Iyengar R.: Suppression of Ras-induced transformation of NIH 3T3 cells by activated G·s. Science. 1994;263:1278-1281.
  • )Vallar L.: Oncogenic role of heterotrimeric G proteins. Cancer Surv. 1996;27:325-338. Nahorski SR.: Pharmacology of intracellular signalling pathways. Br J Pharmacol. 2006; 147:S38-S45.
  • )Sgambati SA, Zarif A, Basson MD.: Octreotide differentially modulates human Caco-2 intestinal epithelial cell proliferation and differentiation by decreasing intracellular cAMP. Regul Pept. 1996; 61: 219-227.
  • )Inoue A, Kuroyanagi Y, Terui K, Moi P, Ikuta T.: Negative regulation of globin gene expression by cyclic AMP-dependent pathway in erythroid cells. Exp Hematol. 2004;32:244-253.
  • )Cokic VP, Andric SA, Stojilkovic SS, Noguchi CT, Schechter AN.: Hydroxyurea nitrosylates and activates soluble guanylyl cyclase in human erythroid cells. Blood. 2008; 111:1117-1123. Küçükkaya B,.Arslan DO, Kan B.: Role of G proteins and ERK activation in hemin-induced erythroid differentiation of K562 cells. Life Sci. 2006;78:1217- 1224.
Toplam 24 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Bölüm Araştırma Makalesi
Yazarlar

Bahire Küçükkaya Bu kişi benim

Yayımlanma Tarihi 1 Ağustos 2010
Yayımlandığı Sayı Yıl 2010 Cilt: 2 Sayı: 2

Kaynak Göster

Vancouver Küçükkaya B. K562 Hücrelerinde Eritroid Farklılaşmayı Uyaran Hemin, Siklik AMP Oluşumunu Baskılar. Maltepe tıp derg. 2010;2(2):32-6.