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Year 2013, Volume: 26 Issue: 2, 72 - 76, 01.10.2015

Hülya Cabadak Banu Aydın Beki Kan

https://doi.org/10.5472/MMJ.2013.02807.1

Abstract

Objectives: Muscarinic acetylcholine receptors (mAChR) belong to a family of G protein coupled receptors (GPCRs). These mAChRs regulate several important physiological functions by activating a wide variety of cellular signaling pathways. We have previously shown that muscarinic acetylcholine (M2, M3 and M4) receptors are expressed in K562 cells. In this study, we investigated the effect of muscarinic agonist, antagonists and different signaling pathway inhibitors on c-Fos and cyclin D1 transcripts, using reverse transcriptase polymerase chain reaction (RT-PCR) that allows changes of very rare transcripts to be monitored. Material and Methods: Total RNA was prepared from K562 cells challenged with muscarinic agonist, antagonists and inhibitors. c-Fos and cyclin D1 expression were determined by RT-PCR.Results: We showed that treatment with muscarinic agonist, antagonists and inhibitors leads to changes in c-Fos and cyclin D1expression in K562 cells.Conclusions: Our results suggest that muscarinic receptors regulate expression of c-Fos and cyclin D1 genes in K562 cells via different signaling pathways

Keywords

Atropine Carbachol (CCh) c-Fos Cyclin D1

References

  • 1. Wessler I, Kilbinger H, Bittinger F, Unger R, Kirkpatrick CJ. The non-neuronal cholinergic system in humans: expression, function and pathophysiology. Life Sci 2003;72:2055–61.doi: 10.1016/S0024- 3205(03)00083-3.
  • 2. Peretto I, Petrillo P, Imbimbo BP. Medicinal chemistry and therapeutic potential of muscarinic M3 antagonists. Med Res Rev 2009;29:867– 902. doi: 10.1002/med.20158.
  • 3. Cabadak H, Aydın B, Kan B. Regulation of M2 , M3 , and M4 muscarinic receptor expression in K562 chronic myelogenous leukemic cells by carbachol. J Recept Signal Transduct Res 2010;31:26-32. doi: 10.3109/10799893.2010.506484.
  • 4. Levey AI. Immunological localization of m1 -m5 muscarinic acethylcholine receptors in peripheral tissues and brain. Life Sci 1993;52:441-8. doi: 10.1016/0024-3205(93)90300-R.
  • 5. Eglen RM, Choppin A, Watson N. Therapeutic opportunities from muscarinic receptor research. Trends Pharmacol Sci 2001;22:409–14. doi: 10.1016/S0165-6147(00)01737-5.
  • 6. Tracey WR, Peach MJ. Differential muscarinic receptor mRNA expression by freshly isolated and cultured bovine aortic endothelial cells. Circ Res 1992;70:234-40. doi: 10.1161/01.RES.70.2.234.
  • 7. Felder CC. Muscarinic acethylcholine receptors: signaltransduction through multiple effectors. FASEB J 1995;9:619–25.
  • 8. Cabadak H, Küçükibrahimoglu E, Aydın B, Kan B,Gören MZ. Muscarinic receptor mediated nitric oxide release in K562 erythroleukemia cell line. Auton Autacoid Pharmacol 2009;29:109–15. doi: 10.1111/j.1474-8673.2009.00431.x
  • 9. Costa LG, Guizzetti M, Oberdoerster J, et al. Modulation of DNA synthesis by muscarinic cholinergic receptors. Growth Factors 2001; 18:227-36. doi: 10.3109/08977190109029ex 112.
  • 10. Harold F, Robert TJ, Dwayne D, Wan-Lin Y and Yinghua X. Human colon cancer cell proliferation mediated by the M3 muscarinic cholinergic receptor1.Clin Cancer Res 1999;5:2532-9.
  • 11. Brown JH, Sah V, Moskowitz S, Ramirez T, Collins L, Post G, Goldstein D. Pathways and roadblocks in muscarinic receptor-me diated growth regulation. Life Sci 1997;60:1077-84.
  • 12. Fujii T. An independent, non-neuronal cholinergic system in lymphocytes and its roles in regulation of immune function. Folia Pharmacol Jpn 2004;123:179-88. doi: 10.1254/fpj.123.179.
  • 13. Fujii T, Kawashima K. Ca2+ oscillation and c-fos gene expression induced via muscarinic acetylcholine receptor in human T- and B-cell lines. Naunyn Schmiedebergs Arch Pharmacol 2000;362:4-21.
  • 14. Ibanez TI, Miwa JM, Wang HL, et al Novel modulation of neuronal nicotinic acethylcholine receptors by association with the endogenous prototoxinlynx1. Neuron 2002;33:893-903. doi: 10.1016/S0896- 6273(02)00632-3.
  • 15. Ding WQ, Larsson C, Alling C. Stimulation of muscarinic receptor induces expression of individual fos and jun genes through different transduction pathways. J Neurochem 1998;70:1722-9.
  • 16. Simonson M S, Jones J M, Dunn M J Differential regulation of cfos and jun gene expression and AP-I cis-element activity by endothelin isopeptides. Possible implications for mitogenic signaling by endothelin. J Biol Chem 1992;267:8643-9.
  • 17. Blackshear PJ, Stumpo DJ, Huang J-K, Nemenoff RA, Spach DH. Protein kinase C-dependent and independent pathways of protooncogene induction in human astrocytoma cells. J Biol Chem 1987;262: 7774-81.
  • 18. Larsson C, Gustavsson L, Simonsson P, Bergman O, Alling C. Mechanisms of muscarinic receptor-stimulated expression of c-fos in SH-SY5Y cells. Euro J Pharmacology 1994;268:19-28. doi: 10.1016/0922-4106(94)90116-3.
  • 19. Fujii T, Kawashima K. Calcium signaling and c-fos gene expression via M3 muscarinic acetylcholine receptors in human T- and B-cells. Jpn J Pharmacol 2000;84:124-32. doi: 10.1254/jjp.84.124.
  • 20. Sato KZ, Fujii T, Watanabe Y, et al Diversity of mRNA expression for muscarinic acetylcholine receptor subtypes and neuronal nicotinic acetylcholine receptor subunits in human mononuclear leukocytes and leukemic cell lines. Neurosci Lett 1999;266:17–20. doi: 10.1016/ S0304-3940(99)00259-1.
  • 21. Cabadak H, Aydın B, Kan B. Muscarinic receptor mediated cAMP response in human K562 chronic myelogenous leukemia cells. Turk J Biochem 2011; 36:188–92.
  • 22. Baldin V, Lukas J, Marcote MJ, Pagano M, Draetta G. Cyclin D1 is a nuclear protein required for cell cycle progression in G1. Genes Dev 1993;7:812-21. doi: 10.1101/gad.7.5.812.
  • 23. Alao JP, Gamble SC, Stavropoulou AV, et al. The cyclin D1 protooncogene is sequestered in the cytoplasm of mammalian cancer cell lines. Mol Cancer 2006;5:7,1-11. doi: 10.1186/1476-4598-5-7.
  • 24. Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987;162:156–9.
  • 25. Jin X, Song X, Li L, et al. Blockade of AP-1 activity by dominantnegative TAM67 can abrogate the oncogenic phenotype in latent membrane protein 1-positive human nasopharyngeal carcinoma. Mol Carcinog 2007;46:901-11. doi: 10.1002/mc.20319.
  • 26. Masuda M, Suzui M, Yasumatu R, et al. Constitutive activation of signal transducers and activators of transcription 3 correlates with cyclin D1 overexpression and may provide a novel prognostic marker in head and neck squamous cell carcinoma. Cancer Res 2002;62:3351–5.
  • 27. Preiksaitis HG, Krysiak PS, Chrones T, Rajgopal V, Laurier LG. Pharmacological and molecular characterization of muscarinic receptor subtypes in human esophageal smooth muscle. J Pharmacol Exp Ther 2000;295:879-88.
  • 28. Caulfield MP, Birdsall NJ. International Union of Pharmacology. XVII. Classification of muscarinic acetylcholine receptors. Pharmacol Rev 1998;50:279–90.
  • 29. Shah N, Khurana S, Cheng K, Raufman JP. Muscarinic receptors and ligands in cancer. Am J Physiol Cell Physiol 2009;296:221-32. doi: 10.1152/ajpcell.00514.2008.
  • 30. Nicke B, Detjen K, Logsdon CD. Muscarinic cholinergic receptors activate both inhibitory and stimulatory growth mechanisms in NIH3T3 cells. J Biol Chem 1999; 274:21701–6. doi: 10.1074/jbc.274.31.21701.
  • 31. Trejo J, Brown JH. c-fos and c-jun are induced by muscarinic receptor activation of protein kinase C but are differentially regulated by intracellular calcium. J Biol Chem 1991;266:7876-82.

K562 hücrelerinde muskarinik agonist, antagonist ve sinyal ileti yolağı inhibitörleri c-Fos ve siklin D1 ekspresyonlarını değiştirir

Year 2013, Volume: 26 Issue: 2, 72 - 76, 01.10.2015

Hülya Cabadak Banu Aydın Beki Kan

https://doi.org/10.5472/MMJ.2013.02807.1

Abstract

Amaç: Muskarinik asetilkolin reseptörleri (mAChR) G protein ile kenetli reseptör ailesinin üyesidirler. mAChR farklı sinyal ileti yolakları aracılığı ile bazı mühim f izyolojik fonksiyonları düzenler. K562 hücrelerinde M2 , M3 ve M4 reseptörlerinin eksprese olduğunu önceki çalışmalarımızda gösterdik. Bu çalışmada muskarinik agonist, antagonist ve farklı sinyal ileti yolağı inhibitörlerinin c-Fos ve siklin D1 ekspresyonuna etkileri araştırılmıştır Gereç ve Yöntem: Muskarinik reseptör agonist, antagonist ve sinyal yolağı inhibitörlerinin c-Fos ve siklin D1 ekspresyonuna etkileri ters transkriptaz polimeraz zincir tepkimesi (TT-PZT) kullanılarak analiz edilmiştir. Bulgular: Muskarinik agonist, antagonistler ve sinyal yolağı inhibitörleri K562 hücrelerinde c-Fos ve siklin D1 transkriptlerinde değişime neden olmuştur. Sonuç: Sonuçlarımız K562 hücrelerinde muskarinik reseptör aracılı c-Fos ve siklin D1 . mRNA ekspresyonlarının farklı sinyal ileti yolları ile düzenlendiğini düşündürmektedir.

Keywords

Atropin c-Fos Karbakol Siklin D1

References

  • 1. Wessler I, Kilbinger H, Bittinger F, Unger R, Kirkpatrick CJ. The non-neuronal cholinergic system in humans: expression, function and pathophysiology. Life Sci 2003;72:2055–61.doi: 10.1016/S0024- 3205(03)00083-3.
  • 2. Peretto I, Petrillo P, Imbimbo BP. Medicinal chemistry and therapeutic potential of muscarinic M3 antagonists. Med Res Rev 2009;29:867– 902. doi: 10.1002/med.20158.
  • 3. Cabadak H, Aydın B, Kan B. Regulation of M2 , M3 , and M4 muscarinic receptor expression in K562 chronic myelogenous leukemic cells by carbachol. J Recept Signal Transduct Res 2010;31:26-32. doi: 10.3109/10799893.2010.506484.
  • 4. Levey AI. Immunological localization of m1 -m5 muscarinic acethylcholine receptors in peripheral tissues and brain. Life Sci 1993;52:441-8. doi: 10.1016/0024-3205(93)90300-R.
  • 5. Eglen RM, Choppin A, Watson N. Therapeutic opportunities from muscarinic receptor research. Trends Pharmacol Sci 2001;22:409–14. doi: 10.1016/S0165-6147(00)01737-5.
  • 6. Tracey WR, Peach MJ. Differential muscarinic receptor mRNA expression by freshly isolated and cultured bovine aortic endothelial cells. Circ Res 1992;70:234-40. doi: 10.1161/01.RES.70.2.234.
  • 7. Felder CC. Muscarinic acethylcholine receptors: signaltransduction through multiple effectors. FASEB J 1995;9:619–25.
  • 8. Cabadak H, Küçükibrahimoglu E, Aydın B, Kan B,Gören MZ. Muscarinic receptor mediated nitric oxide release in K562 erythroleukemia cell line. Auton Autacoid Pharmacol 2009;29:109–15. doi: 10.1111/j.1474-8673.2009.00431.x
  • 9. Costa LG, Guizzetti M, Oberdoerster J, et al. Modulation of DNA synthesis by muscarinic cholinergic receptors. Growth Factors 2001; 18:227-36. doi: 10.3109/08977190109029ex 112.
  • 10. Harold F, Robert TJ, Dwayne D, Wan-Lin Y and Yinghua X. Human colon cancer cell proliferation mediated by the M3 muscarinic cholinergic receptor1.Clin Cancer Res 1999;5:2532-9.
  • 11. Brown JH, Sah V, Moskowitz S, Ramirez T, Collins L, Post G, Goldstein D. Pathways and roadblocks in muscarinic receptor-me diated growth regulation. Life Sci 1997;60:1077-84.
  • 12. Fujii T. An independent, non-neuronal cholinergic system in lymphocytes and its roles in regulation of immune function. Folia Pharmacol Jpn 2004;123:179-88. doi: 10.1254/fpj.123.179.
  • 13. Fujii T, Kawashima K. Ca2+ oscillation and c-fos gene expression induced via muscarinic acetylcholine receptor in human T- and B-cell lines. Naunyn Schmiedebergs Arch Pharmacol 2000;362:4-21.
  • 14. Ibanez TI, Miwa JM, Wang HL, et al Novel modulation of neuronal nicotinic acethylcholine receptors by association with the endogenous prototoxinlynx1. Neuron 2002;33:893-903. doi: 10.1016/S0896- 6273(02)00632-3.
  • 15. Ding WQ, Larsson C, Alling C. Stimulation of muscarinic receptor induces expression of individual fos and jun genes through different transduction pathways. J Neurochem 1998;70:1722-9.
  • 16. Simonson M S, Jones J M, Dunn M J Differential regulation of cfos and jun gene expression and AP-I cis-element activity by endothelin isopeptides. Possible implications for mitogenic signaling by endothelin. J Biol Chem 1992;267:8643-9.
  • 17. Blackshear PJ, Stumpo DJ, Huang J-K, Nemenoff RA, Spach DH. Protein kinase C-dependent and independent pathways of protooncogene induction in human astrocytoma cells. J Biol Chem 1987;262: 7774-81.
  • 18. Larsson C, Gustavsson L, Simonsson P, Bergman O, Alling C. Mechanisms of muscarinic receptor-stimulated expression of c-fos in SH-SY5Y cells. Euro J Pharmacology 1994;268:19-28. doi: 10.1016/0922-4106(94)90116-3.
  • 19. Fujii T, Kawashima K. Calcium signaling and c-fos gene expression via M3 muscarinic acetylcholine receptors in human T- and B-cells. Jpn J Pharmacol 2000;84:124-32. doi: 10.1254/jjp.84.124.
  • 20. Sato KZ, Fujii T, Watanabe Y, et al Diversity of mRNA expression for muscarinic acetylcholine receptor subtypes and neuronal nicotinic acetylcholine receptor subunits in human mononuclear leukocytes and leukemic cell lines. Neurosci Lett 1999;266:17–20. doi: 10.1016/ S0304-3940(99)00259-1.
  • 21. Cabadak H, Aydın B, Kan B. Muscarinic receptor mediated cAMP response in human K562 chronic myelogenous leukemia cells. Turk J Biochem 2011; 36:188–92.
  • 22. Baldin V, Lukas J, Marcote MJ, Pagano M, Draetta G. Cyclin D1 is a nuclear protein required for cell cycle progression in G1. Genes Dev 1993;7:812-21. doi: 10.1101/gad.7.5.812.
  • 23. Alao JP, Gamble SC, Stavropoulou AV, et al. The cyclin D1 protooncogene is sequestered in the cytoplasm of mammalian cancer cell lines. Mol Cancer 2006;5:7,1-11. doi: 10.1186/1476-4598-5-7.
  • 24. Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987;162:156–9.
  • 25. Jin X, Song X, Li L, et al. Blockade of AP-1 activity by dominantnegative TAM67 can abrogate the oncogenic phenotype in latent membrane protein 1-positive human nasopharyngeal carcinoma. Mol Carcinog 2007;46:901-11. doi: 10.1002/mc.20319.
  • 26. Masuda M, Suzui M, Yasumatu R, et al. Constitutive activation of signal transducers and activators of transcription 3 correlates with cyclin D1 overexpression and may provide a novel prognostic marker in head and neck squamous cell carcinoma. Cancer Res 2002;62:3351–5.
  • 27. Preiksaitis HG, Krysiak PS, Chrones T, Rajgopal V, Laurier LG. Pharmacological and molecular characterization of muscarinic receptor subtypes in human esophageal smooth muscle. J Pharmacol Exp Ther 2000;295:879-88.
  • 28. Caulfield MP, Birdsall NJ. International Union of Pharmacology. XVII. Classification of muscarinic acetylcholine receptors. Pharmacol Rev 1998;50:279–90.
  • 29. Shah N, Khurana S, Cheng K, Raufman JP. Muscarinic receptors and ligands in cancer. Am J Physiol Cell Physiol 2009;296:221-32. doi: 10.1152/ajpcell.00514.2008.
  • 30. Nicke B, Detjen K, Logsdon CD. Muscarinic cholinergic receptors activate both inhibitory and stimulatory growth mechanisms in NIH3T3 cells. J Biol Chem 1999; 274:21701–6. doi: 10.1074/jbc.274.31.21701.
  • 31. Trejo J, Brown JH. c-fos and c-jun are induced by muscarinic receptor activation of protein kinase C but are differentially regulated by intracellular calcium. J Biol Chem 1991;266:7876-82.
There are 31 citations in total.

Details

Primary Language Turkish
Journal Section Articles
Authors

Hülya Cabadak This is me

Banu Aydın This is me

Beki Kan This is me

Publication Date October 1, 2015
Published in Issue Year 2013 Volume: 26 Issue: 2

Cite

APA Cabadak, H., Aydın, B., & Kan, B. (2015). K562 hücrelerinde muskarinik agonist, antagonist ve sinyal ileti yolağı inhibitörleri c-Fos ve siklin D1 ekspresyonlarını değiştirir. Marmara Medical Journal, 26(2), 72-76. https://doi.org/10.5472/MMJ.2013.02807.1
AMA Cabadak H, Aydın B, Kan B. K562 hücrelerinde muskarinik agonist, antagonist ve sinyal ileti yolağı inhibitörleri c-Fos ve siklin D1 ekspresyonlarını değiştirir. Marmara Med J. October 2015;26(2):72-76. doi:10.5472/MMJ.2013.02807.1
Chicago Cabadak, Hülya, Banu Aydın, and Beki Kan. “K562 hücrelerinde Muskarinik Agonist, Antagonist Ve Sinyal Ileti yolağı inhibitörleri C-Fos Ve Siklin D1 ekspresyonlarını değiştirir”. Marmara Medical Journal 26, no. 2 (October 2015): 72-76. https://doi.org/10.5472/MMJ.2013.02807.1.
EndNote Cabadak H, Aydın B, Kan B (October 1, 2015) K562 hücrelerinde muskarinik agonist, antagonist ve sinyal ileti yolağı inhibitörleri c-Fos ve siklin D1 ekspresyonlarını değiştirir. Marmara Medical Journal 26 2 72–76.
IEEE H. Cabadak, B. Aydın, and B. Kan, “K562 hücrelerinde muskarinik agonist, antagonist ve sinyal ileti yolağı inhibitörleri c-Fos ve siklin D1 ekspresyonlarını değiştirir”, Marmara Med J, vol. 26, no. 2, pp. 72–76, 2015, doi: 10.5472/MMJ.2013.02807.1.
ISNAD Cabadak, Hülya et al. “K562 hücrelerinde Muskarinik Agonist, Antagonist Ve Sinyal Ileti yolağı inhibitörleri C-Fos Ve Siklin D1 ekspresyonlarını değiştirir”. Marmara Medical Journal 26/2 (October 2015), 72-76. https://doi.org/10.5472/MMJ.2013.02807.1.
JAMA Cabadak H, Aydın B, Kan B. K562 hücrelerinde muskarinik agonist, antagonist ve sinyal ileti yolağı inhibitörleri c-Fos ve siklin D1 ekspresyonlarını değiştirir. Marmara Med J. 2015;26:72–76.
MLA Cabadak, Hülya et al. “K562 hücrelerinde Muskarinik Agonist, Antagonist Ve Sinyal Ileti yolağı inhibitörleri C-Fos Ve Siklin D1 ekspresyonlarını değiştirir”. Marmara Medical Journal, vol. 26, no. 2, 2015, pp. 72-76, doi:10.5472/MMJ.2013.02807.1.
Vancouver Cabadak H, Aydın B, Kan B. K562 hücrelerinde muskarinik agonist, antagonist ve sinyal ileti yolağı inhibitörleri c-Fos ve siklin D1 ekspresyonlarını değiştirir. Marmara Med J. 2015;26(2):72-6.