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Effects of cholinergic compounds and TNF-alpha on human erythroleukemia K562 cell proliferation and caspase expression

Yıl 2019, , 20 - 26, 29.01.2019
https://doi.org/10.5472/marumj.518797

Öz

Objective: The purpose of this study was to investigate if
stimulating auto-paracrine muscarinic receptor signalling pathway
could change human erythroleukemia K562 cell proliferation and
caspase 3, 8 and 9 expression levels. To better understand the role of
muscarinic receptors in cell signalling mechanism, we investigated
the effects of several compounds on human erythroleukemia
K562 cell proliferation and caspase 3, 8 and 9 expression. These
compounds were M3 muscarinic receptor agonist, pilocarpine, proinflammatory
cytokine, tumor necrosis factor (TNF)-alpha, and
the wortmannin which is a phosphoinositide 3-kinase inhibitor.
Materials and Methods: Cell proliferation and cell viability
were evaluated by the trypan blue exclusion test and 5-Bromo-2-
deoxy-uridine (BrdU) Labelling and Detection Kits. Caspase 3, 8
and 9 expression levels were determined by immunoblot analysis.
Results: Both pilocarpine and TNF-alpha caused a small increase
in human erythroleukemia K562 cell proliferation. However, when
all the compounds were treated together, proliferation of human
erythroleukemia K562 cells increased significantly when compared to
untreated control cells. TNF-alpha and wortmannin treatment increased
caspase 3 and caspase 8 expression patterns significantly in human
erythroleukemia K562 cells. TNF-alpha and wortmannin treatment
increased caspase 9 expression level (P>0.05) but it was not significant.
Conclusion: These findings partly demonstrated that M3
muscarinic receptor mediated an increase in K562 cell proliferation.
Pilocarpine prevented TNF-alpha and wortmannin induced
caspase 3 and 8 expression and indirectly showed apoptosis in
human erythroleukemia K562 cells.

Kaynakça

  • 1. Tong Y, Liu YY, You LS, et al. Perifosine induces protective autophagy and upregulation of ATG5 in human chronic myelogenous leukemia cells in vitro. Acta Pharmacol Sin 2012; 33: 542-50. doi.org/ 10.1038/aps.2011.192
  • 2. Lozzio CB, Lozzio BB. Human chronic myelogenous leukemia cell line with positive Philadelphia chromosome. Blood 1975; 45: 321-34. doi.org/10.1182/ blood-2016-08-736025
  • 3. Bonner TI, Buckley NJ, Young AC, et al. Identification of a family of muscarinic acetylcholine receptor genes. Science 1987; 237:527-32. doi.org/10.1126 /science .3037705
  • 4. Kawashima K, Fujii T. Basic and clinical aspects of non-neuronal acetylcholine: Overview of non-neuronal cholinergic systems and their biological significance. J Pharmacol Sci 2008; 106: 167-73. doi.org/ 10.1254/jphs. FM0070073
  • 5. Felder CC. Muscarinic acetylcholine receptors: Signal transduction through multiple effectors. FASEB J 1995; 9:619-25. doi: 10.1096/fasebj.9.8.7768353.
  • 6. Guizzetti M, Costa LG. Activation of phosphatidylinositol 3kinase by muscarinic receptors in astrocytoma cells. Neuroreport 2001; 12: 1639-42. doi: 10.1097/00001.756.200106130-00025
  • 7. Cabadak H, Kücükibrahimoğlu E, Aydin B, ark. Muscarinic receptor-mediated nitric oxide release in a K562 erythroleukaemia cell line. Auton Autacoid Pharmacol 2009; 29: 109-15. doi.org/ 10.1111/j.I474-8673.2009.0043.x
  • 8. Cabadak H, Aydin B, Kan B. Regulation of M2, M3, and M4 muscarinic receptor expression in K562 chronic myelogenous leukemic cells by carbachol. J Recept Signal Transd Res 2011; 31:26-32. doi.org/ 10.3109/10799.893.2010.506484
  • 9. Aydın B, Kan B, Cabadak H. The role of intracellular pathways in the proliferation of human K562 cells that is mediated by muscarinic receptors. Leukemia Res 2013; 37:1144-9. doi.org/10.1016/j.leukres.2013.05.018
  • 10. Fadeel B, Orrenius S. Apoptosis: a basic biological phenomenon with wide-ranging implications in human disease. J Intern Med 2005; 258:479-517. doi.org/10.1111 /j. 1365-2796.2005.01570.x
  • 11. Igney FH, Krammer PH. Death and anti-death: tumor resistance to apoptosis. Nat Rev Cancer 2002; 2:277-88. doi. org/10.1038/nrc776
  • 12. Baud V,Karin M. Signal transduction by tumor necrosis factor and its relatives. Trends Cell Biol 2001; 9:372-7. doi. org/ 10.1016/S0962-8924(01)02064-5
  • 13. Boatright KM, Salvesen GS. Mechanisms of caspase activation. Curr Opin Cell Biol 2003; 15:725-31. doi. org/10.1016/j.ceb.2003.10.009
  • 14. Ware CF, VanArsdale S, VanArsdale TL. Apoptosis mediated by the TNF-alpha related cytokine and receptor families. J Cell Biol 1996; 60:47-55. doi.org/10.1002 10.1002/(SICI)1097-4644(19960101)60:1%3C47::AIDJCB8% 3E3.0.CO;2-3
  • 15. Lowry OH, Rosebrough NJ, Farr AL, et al. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-75.
  • 16. Eliot R. Spindel. Muscarinic receptor agonists and antagonists: Effects on Cancer. Handb Exp Pharmacol 2012; 208: 451-468. doi.org/ 10.1007/978-3-642-23274-9-19
  • 17. Goeddel DV, Aggarwal BB, Gray PW, et al. Tumor necrosis factors: gene structure and biological activities. Cold Spring Harb Symp Quant Biol 51, 1986: 597-609. doi.org/ 10.1101/ SQB.1986.051.01.072
  • 18. Gonzalez-Flores D, Rodrignez AB, Pariente JA. TNFalpha- induced apoptosis in human myeloid cell lines HL- 60 and K562 is dependent of intracellular ROS generation. Mol Cellular Biochem 2014; 390:281-7. doi.org/ 10.1007/ s11010.014.1979-5
  • 19. Xuan P, Storr SJ, Zhang Y, et al. Caspase-3 and caspase-8 expression in breast cancer: caspase-3 is associated with survival. Apoptosis 2017; 22: 357-68. doi.org/ 10.1016/ S0898-6568(02)00025-6
  • 20. Corda S, Laplace C, Vicaut E, et al. Rapid reactive oxygen species production by mitochondria in endothelial cells exposed to tumor necrosis factor-alpha is mediated by ceramide. Am J Respir Cell Mol Biol 2001; 24: 762-8. doi. Org/ 10.1080/15384.047.2015.1029835
  • 21. Sandra F, Matsuki NA, Takeuchi H, et al. TNF inhibited the apoptosis by activation of Akt serine/threonine kinase in the human head and neck squamous cell carcinoma. Cell Signal 2002; 14:771-8. doi.org/10.1016/ S0898-6568(02)00025-6
  • 22. Olsen BB, Bjørling-Poulsen M, Guerra B. Emodin negatively affects the phosphoinositide 3-kinase/AKT signalling pathway: a study on its mechanism of action. Int J Biochem Cell Biol 2007; 39:227-37. doi:10.1016/j.biocel.2006.08. 006
  • 23. Rudelius M, Pittaluga S, Nishizuka S, et al. Constitutive activation of Akt contributes to the pathogenesis and survival of mantle cell lymphoma. Blood 2006; 108:1668-1676. doi:10.1182/blood-2006-04-015586
  • 24. Wu Q, Chen Y, Cui G, Cheng Y. Wortmannin inhibits K562 leukemic cells by regulating PI3k/Akt channel in vitro. J Huazhong Univ Sci Technolog Med Sci. 2009; 29:451-6. doi: 10.1007/s11596.009.0412-x
  • 25. Zhao Q, Gu X, Zhang C, et al. Blocking M2 muscarinic receptor signaling inhibits tumor growth and reverses epithelial-mesenchymal transition (EMT) in non-small cell lung cancer (NSCLC). Cancer Biol Ther 2015; 16:634-43. doi:10.1080 /153840 47.2015.102.9835

Kolinerjik bileşiklerin ve TNF-alfanın insan eritrolösemi K562 hücre çoğalmasına ve kaspaz ekspresyonu üzerine etkileri

Yıl 2019, , 20 - 26, 29.01.2019
https://doi.org/10.5472/marumj.518797

Öz

Amaç: Bu çalışmanın amacı, otoparakrin M3 muskarinik
reseptör sinyal yolağının uyarılmasının, insan eritrolösemi K562
hücrelerinin çoğalmasında ve kaspaz 3, 8 ve 9 ekspresyon seviyeleri
üzerinde etkisi olup olmadığını araştırmaktır. Hücre sinyal
ileti mekanizmasında muskarinik reseptörlerin rolünü daha iyi
anlamak üzere, çeşitli bileşiklerin insan eritrolösemisi K562 hücre
proliferasyonu ve kaspaz 3, 8 ve 9 ekspresyon seviyeleri üzerindeki
etkilerini araştırdık. Bu bileşikler, M3 muskarinik reseptör agonisti,
pilokarpin, pro-enflamatuvar sitokin, tümör nekroz faktör (TNF)-
alfa ve fosfoinositid 3-kinaz inhibitörü wortmanindir.
Gereçler ve Yöntemler: Hücre çoğalması ve hücre canlılığı,
tripan mavisi testi ve 5-Bromo-2-deoxy-uridine (BrdU) İşaretleme
ve Belirleme kitleri ile değerlendirildi. Kaspaz 3, 8 ve 9 ekspresyon
seviyeleri immunoblot analizi ile belirlendi.
Bulgular: Hem pilokarpin, hem de TNF-alfa, insan eritrolösemi
K562 hücre çoğalmasında çok az artışa neden oldu. Ancak, tüm
bileşikler birlikte muamele edildiğinde, insan eritrolösemi K562
hücrelerinin çoğalması, muamele edilmeyen kontrol hücrelerine
göre anlamlı olarak arttı. TNF-alfa ve wortmanin ile muamele
edilen K562 hücrelerinde kaspaz 3 ve kaspaz 8 ekspresyonu
seviyelerinde anlamlı değişim belirlendi. TNF-alfa ve wortmannin
muamelesi kaspaz 9 ekspresyon seviyesini arttırdı (P> 0.05) ancak
anlamlı değildi.
Sonuç: Bu bulgular, kısmen M3 muskarinik reseptör aracılı
K562 hücre çoğalmasında artış olduğunu göstermektedir.
Pilokarpin, insan eritrolösemi K562 hücrelerinde, TNF-alfa ve
wortmanin ile uyarılan kaspaz 3 ve 8 ekspresyonunu önledi ve
dolaylı olarak apoptozu gösterdi.

Kaynakça

  • 1. Tong Y, Liu YY, You LS, et al. Perifosine induces protective autophagy and upregulation of ATG5 in human chronic myelogenous leukemia cells in vitro. Acta Pharmacol Sin 2012; 33: 542-50. doi.org/ 10.1038/aps.2011.192
  • 2. Lozzio CB, Lozzio BB. Human chronic myelogenous leukemia cell line with positive Philadelphia chromosome. Blood 1975; 45: 321-34. doi.org/10.1182/ blood-2016-08-736025
  • 3. Bonner TI, Buckley NJ, Young AC, et al. Identification of a family of muscarinic acetylcholine receptor genes. Science 1987; 237:527-32. doi.org/10.1126 /science .3037705
  • 4. Kawashima K, Fujii T. Basic and clinical aspects of non-neuronal acetylcholine: Overview of non-neuronal cholinergic systems and their biological significance. J Pharmacol Sci 2008; 106: 167-73. doi.org/ 10.1254/jphs. FM0070073
  • 5. Felder CC. Muscarinic acetylcholine receptors: Signal transduction through multiple effectors. FASEB J 1995; 9:619-25. doi: 10.1096/fasebj.9.8.7768353.
  • 6. Guizzetti M, Costa LG. Activation of phosphatidylinositol 3kinase by muscarinic receptors in astrocytoma cells. Neuroreport 2001; 12: 1639-42. doi: 10.1097/00001.756.200106130-00025
  • 7. Cabadak H, Kücükibrahimoğlu E, Aydin B, ark. Muscarinic receptor-mediated nitric oxide release in a K562 erythroleukaemia cell line. Auton Autacoid Pharmacol 2009; 29: 109-15. doi.org/ 10.1111/j.I474-8673.2009.0043.x
  • 8. Cabadak H, Aydin B, Kan B. Regulation of M2, M3, and M4 muscarinic receptor expression in K562 chronic myelogenous leukemic cells by carbachol. J Recept Signal Transd Res 2011; 31:26-32. doi.org/ 10.3109/10799.893.2010.506484
  • 9. Aydın B, Kan B, Cabadak H. The role of intracellular pathways in the proliferation of human K562 cells that is mediated by muscarinic receptors. Leukemia Res 2013; 37:1144-9. doi.org/10.1016/j.leukres.2013.05.018
  • 10. Fadeel B, Orrenius S. Apoptosis: a basic biological phenomenon with wide-ranging implications in human disease. J Intern Med 2005; 258:479-517. doi.org/10.1111 /j. 1365-2796.2005.01570.x
  • 11. Igney FH, Krammer PH. Death and anti-death: tumor resistance to apoptosis. Nat Rev Cancer 2002; 2:277-88. doi. org/10.1038/nrc776
  • 12. Baud V,Karin M. Signal transduction by tumor necrosis factor and its relatives. Trends Cell Biol 2001; 9:372-7. doi. org/ 10.1016/S0962-8924(01)02064-5
  • 13. Boatright KM, Salvesen GS. Mechanisms of caspase activation. Curr Opin Cell Biol 2003; 15:725-31. doi. org/10.1016/j.ceb.2003.10.009
  • 14. Ware CF, VanArsdale S, VanArsdale TL. Apoptosis mediated by the TNF-alpha related cytokine and receptor families. J Cell Biol 1996; 60:47-55. doi.org/10.1002 10.1002/(SICI)1097-4644(19960101)60:1%3C47::AIDJCB8% 3E3.0.CO;2-3
  • 15. Lowry OH, Rosebrough NJ, Farr AL, et al. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-75.
  • 16. Eliot R. Spindel. Muscarinic receptor agonists and antagonists: Effects on Cancer. Handb Exp Pharmacol 2012; 208: 451-468. doi.org/ 10.1007/978-3-642-23274-9-19
  • 17. Goeddel DV, Aggarwal BB, Gray PW, et al. Tumor necrosis factors: gene structure and biological activities. Cold Spring Harb Symp Quant Biol 51, 1986: 597-609. doi.org/ 10.1101/ SQB.1986.051.01.072
  • 18. Gonzalez-Flores D, Rodrignez AB, Pariente JA. TNFalpha- induced apoptosis in human myeloid cell lines HL- 60 and K562 is dependent of intracellular ROS generation. Mol Cellular Biochem 2014; 390:281-7. doi.org/ 10.1007/ s11010.014.1979-5
  • 19. Xuan P, Storr SJ, Zhang Y, et al. Caspase-3 and caspase-8 expression in breast cancer: caspase-3 is associated with survival. Apoptosis 2017; 22: 357-68. doi.org/ 10.1016/ S0898-6568(02)00025-6
  • 20. Corda S, Laplace C, Vicaut E, et al. Rapid reactive oxygen species production by mitochondria in endothelial cells exposed to tumor necrosis factor-alpha is mediated by ceramide. Am J Respir Cell Mol Biol 2001; 24: 762-8. doi. Org/ 10.1080/15384.047.2015.1029835
  • 21. Sandra F, Matsuki NA, Takeuchi H, et al. TNF inhibited the apoptosis by activation of Akt serine/threonine kinase in the human head and neck squamous cell carcinoma. Cell Signal 2002; 14:771-8. doi.org/10.1016/ S0898-6568(02)00025-6
  • 22. Olsen BB, Bjørling-Poulsen M, Guerra B. Emodin negatively affects the phosphoinositide 3-kinase/AKT signalling pathway: a study on its mechanism of action. Int J Biochem Cell Biol 2007; 39:227-37. doi:10.1016/j.biocel.2006.08. 006
  • 23. Rudelius M, Pittaluga S, Nishizuka S, et al. Constitutive activation of Akt contributes to the pathogenesis and survival of mantle cell lymphoma. Blood 2006; 108:1668-1676. doi:10.1182/blood-2006-04-015586
  • 24. Wu Q, Chen Y, Cui G, Cheng Y. Wortmannin inhibits K562 leukemic cells by regulating PI3k/Akt channel in vitro. J Huazhong Univ Sci Technolog Med Sci. 2009; 29:451-6. doi: 10.1007/s11596.009.0412-x
  • 25. Zhao Q, Gu X, Zhang C, et al. Blocking M2 muscarinic receptor signaling inhibits tumor growth and reverses epithelial-mesenchymal transition (EMT) in non-small cell lung cancer (NSCLC). Cancer Biol Ther 2015; 16:634-43. doi:10.1080 /153840 47.2015.102.9835
Toplam 25 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Klinik Tıp Bilimleri
Bölüm Makaleler
Yazarlar

Zehra Kanlı Bu kişi benim

Banu Aydın Bu kişi benim

Hulya Cabadak Bu kişi benim

Yayımlanma Tarihi 29 Ocak 2019
Yayımlandığı Sayı Yıl 2019

Kaynak Göster

APA Kanlı, Z., Aydın, B., & Cabadak, H. (2019). Effects of cholinergic compounds and TNF-alpha on human erythroleukemia K562 cell proliferation and caspase expression. Marmara Medical Journal, 32(1), 20-26. https://doi.org/10.5472/marumj.518797
AMA Kanlı Z, Aydın B, Cabadak H. Effects of cholinergic compounds and TNF-alpha on human erythroleukemia K562 cell proliferation and caspase expression. Marmara Med J. Ocak 2019;32(1):20-26. doi:10.5472/marumj.518797
Chicago Kanlı, Zehra, Banu Aydın, ve Hulya Cabadak. “Effects of Cholinergic Compounds and TNF-Alpha on Human Erythroleukemia K562 Cell Proliferation and Caspase Expression”. Marmara Medical Journal 32, sy. 1 (Ocak 2019): 20-26. https://doi.org/10.5472/marumj.518797.
EndNote Kanlı Z, Aydın B, Cabadak H (01 Ocak 2019) Effects of cholinergic compounds and TNF-alpha on human erythroleukemia K562 cell proliferation and caspase expression. Marmara Medical Journal 32 1 20–26.
IEEE Z. Kanlı, B. Aydın, ve H. Cabadak, “Effects of cholinergic compounds and TNF-alpha on human erythroleukemia K562 cell proliferation and caspase expression”, Marmara Med J, c. 32, sy. 1, ss. 20–26, 2019, doi: 10.5472/marumj.518797.
ISNAD Kanlı, Zehra vd. “Effects of Cholinergic Compounds and TNF-Alpha on Human Erythroleukemia K562 Cell Proliferation and Caspase Expression”. Marmara Medical Journal 32/1 (Ocak 2019), 20-26. https://doi.org/10.5472/marumj.518797.
JAMA Kanlı Z, Aydın B, Cabadak H. Effects of cholinergic compounds and TNF-alpha on human erythroleukemia K562 cell proliferation and caspase expression. Marmara Med J. 2019;32:20–26.
MLA Kanlı, Zehra vd. “Effects of Cholinergic Compounds and TNF-Alpha on Human Erythroleukemia K562 Cell Proliferation and Caspase Expression”. Marmara Medical Journal, c. 32, sy. 1, 2019, ss. 20-26, doi:10.5472/marumj.518797.
Vancouver Kanlı Z, Aydın B, Cabadak H. Effects of cholinergic compounds and TNF-alpha on human erythroleukemia K562 cell proliferation and caspase expression. Marmara Med J. 2019;32(1):20-6.