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NRK-52E Hücre Serisinde Timokinon İndüklü PI3K/AKT/mTOR Yolak Aktivasyonu

Year 2021, Volume: 11 Issue: 1, 68 - 74, 01.03.2021
https://doi.org/10.21597/jist.817666

Abstract

Çalışma birçok hastalığın geleneksel tedavisinde kullanılan Nigella sativa (çörek otu)’nın önemli içeriliklerinden biri olan ve bazı ilaçların üretiminde kullanılan timokinonun (TQ), hücre proliferasyonunda rol oynayan PI3K/AKT/mTOR yolağında yer alan kavşak genlerinin ekspresyon seviyesine etkisini araştırmak amacıyla gerçekleştirildi. Çalışma materyali olarak NRK-52E hücre serisi kullanıldı. Hücrelere farklı konsantrasyonlarda TQ uygulanarak MTT canlılık testi ile TQ nun proliferatif konsantrasyonu belirlendi. Belirlenen TQ konsantrasyonu hücrelere uygulandı ve PI3K/AKT/mTOR yolağında yer alan önemli kavşak genlerin ekspresyon düzeyleri gerçek zamanlı kantitatif polimeraz zincir reaksiyonu (RT-qPCR) yöntemiyle tespit edildi. TQ’nun belli konsantrasyona kadar hücre canlılığını artırdığı, sonrasında ise artan konsantrasyon miktarıyla birlikte sitotoksite oluşturduğu tespit edildi. TQ’nun proliferatif konsantrasyonu 10 µM olarak belirlendi. TQ uygulandıktan 24 saat sonra bir reseptör kinaz alt ünitesi olan ERBB2 artışı ile birlikte PI3K ve AKT1 gen düzeylerinin de arttığı, buna karşın mTOR ekspresyon düzeyinde azalma olduğu belirlendi. Bu verilere göre, geleneksel tıpta çokça kullanılan ve içeriğinde timokinon bulunan N. Sativa’nın düşük konsantrasyonda tüketilmesinin yararlı olabileceği, buna karşın yüksek konsantrasyonda tüketilmesinin böbrek hasarına yol açabileceği düşünülmektedir. Bu olasılığının araştırılmaya değer olduğu ve bu amaçla daha ileri çalışmaların planlanmasının yararlı olabileceği, TQ’un moleküler düzeyde farklı hücre türlerine olan etkisinin araştırılmasında ileriki çalışmalara ışık tutabileceği sonucuna varılmıştır.

References

  • Bai T, Lian LH, Wu YL, Wan Y, Nan JX, 2013. Thymoquinone Attenuates Liver Fibrosis via PI3K and TLR4 Signaling Pathways in Activated Hepatic Stellate Cells. International Immunopharmacology, 15(2):275-281. doi: 10.1016/j.intimp.2012.12.020.
  • Beck JT, Ismail A, Tolomeo C, 2014. Targeting The Phosphatidylinositol 3-kinase (PI3K)/AKT/Mammalian Target of Rapamycin (mTOR) Pathway: an Emerging Treatment Strategy for Squamous Cell Lung Carcinoma. Cancer Treatment Reviews, 40(8): 980-989. doi: 10.1016/j.ctrv.2014.06.006.
  • Brazil DP, Hemmings BA, 2001. Ten Years of Protein Kinase B Signalling: A Hard Akt to Follow. Trends in Biochemical Sciences, 26(11): 657-664. doi: 10.1016/s0968-0004(01)01958-2.
  • Dede S, Yur F, Taşpınar M, Çetin S, Usta A, Yüksek V, 2019. In Vitro Evaluation of Thymoquinone and Lycopene Supplementation on Oxidative DNA Damage and Oxidant Status in High Glucose Conditions. Latin American Journal Pharmacy, 38 (1): 209-912.
  • Ghosheh OA, Houdi A, Crooks PA, 1999. High Performance Liquid Chromatography Analysis of The Pharmacologically Active Quinines and Related Compounds in The Oil of The Black Seed (Nigella Sativa). Journal of Pharmaceutical and Biomedical Analysis, 19(5): 757-762.
  • Hemmings BA, Restuccia DF, 2015. PI3K-PKB/Akt Pathway. Cold Spring Harbor Perspectives in Biology, 1;4(9): a011189. doi: 10.1101/cshperspect.a011189.
  • Hosseinzadeh H, Taiari S, Nassiri-Asl M, 2012. Effect of Thymoquinone, a Constituent of Nigella Sativa L., On İschemia–Reperfusion İn Rat Skeletal Muscle. Naunyn-Schmiedeberg's Archives of Pharmacology, 385(5), 503-508.
  • Iskender B, Izgi K, Canatan H, 2016. Novel Anti-Cancer Agent Myrtucommulone-A And Thymoquinone Abrogate Epithelial-Mesenchymal Transition in Cancer Cells Mainly Through The İnhibition of PI3K/AKT Signalling Axis. Molecular and Cellular Biochemistry, 416(1-2):71-84. doi: 10.1007/s11010-016-2697-y.
  • Khader M, Eckl PM, 2014. Thymoquinone: An Emerging Natural Drug withth a Wide Range of Medical Applications. Iran Journal Basic Medicine Science, 17(12): 950-957
  • Porta C, Paglino C, Mosca A, 2014. Targeting PI3K/Akt/mTOR Signaling in Cancer. Frontiers in Oncology, 14;4:64. doi: 10.3389/fonc.2014.00064.
  • Xu D, Ma Y, Zhao B, Li S, Zhang Y, Pan S, Wu Y, Wang J, Wang D, Pan H, Liu L, Jiang H, 2014. Thymoquinone Induces G2/M Arrest, Inactivates PI3K/Akt and Nuclear factor-κB Pathways in Human Cholangiocarcinomas both in vitro and in vivo. Oncology Reports, 31(5):2063-2070. doi: 10.3892/or.2014.3059..
  • Yang XL, Lin FJ, Guo YJ, Shao ZM, Ou ZL, 2014. Gemcitabine Resistance in Breast Cancer Cells Regulated By PI3K/AKT-Mediated Cellular Proliferation Exerts Negative Feedback via The MEK/MAPK and mTOR Pathways. OncoTargets and Therapy, 7:1033-1042. doi: 10.2147/OTT.S63145.
  • Yüksek V, Dede S, Taşpınar M, Yur F, Çetin S, 2017. Effects of Thymoquınone Applicatıon in The Renal Cell Line. International Congress on Medicinal and Aromatic Plants, Konya, May 10-12, 2017, pp: 842-844.

Activation of PI3K/AKT/mTOR Pathway Thymoquinone-induced in NRK-52E Cell Line

Year 2021, Volume: 11 Issue: 1, 68 - 74, 01.03.2021
https://doi.org/10.21597/jist.817666

Abstract

The study was carried out to investigate the effect of thymoquinone (TQ), one of the important ingredients of Nigella sativa (N. Sativa-black seed) used in traditional treatment of many diseases and production of some drugs, on the expression level of some major genes involved in cell proliferation in the PI3K/AKT/mTOR pathway. NRK-52E cell line was used as study material. The proliferative concentration of TQ was determined by MTT viability assay by treating different concentrations of TQ on the cells. The determined TQ concentration was administered to the cells and the expression levels of the important junctional genes in the PI3K/AKT/mTOR pathway were determined by real-time quantitative polymerase chain reaction (RT-qPCR). It was determined that TQ increased cell viability up to a certain concentration, and then caused cytotoxicity with increasing concentration. In this study, the proliferative concentration of TQ was determined as 10 µM. It was detected that 24 hours after TQ administration, ERBB2, a receptor tyrosine kinase subunit, increased with the increase in PI3K and AKT1 gene levels, however a decrease in the mTOR expression level. According to these data, it is thought that consuming TQ and its contents in low concentrations may be beneficial, whereas consumption in high concentrations may lead to kidney damage. It was concluded that this possibility is worth investigating and it may be useful to plan further studies for this purpose and shed light on future studies in investigating the effect of TQ on different cell types at the molecular level.

References

  • Bai T, Lian LH, Wu YL, Wan Y, Nan JX, 2013. Thymoquinone Attenuates Liver Fibrosis via PI3K and TLR4 Signaling Pathways in Activated Hepatic Stellate Cells. International Immunopharmacology, 15(2):275-281. doi: 10.1016/j.intimp.2012.12.020.
  • Beck JT, Ismail A, Tolomeo C, 2014. Targeting The Phosphatidylinositol 3-kinase (PI3K)/AKT/Mammalian Target of Rapamycin (mTOR) Pathway: an Emerging Treatment Strategy for Squamous Cell Lung Carcinoma. Cancer Treatment Reviews, 40(8): 980-989. doi: 10.1016/j.ctrv.2014.06.006.
  • Brazil DP, Hemmings BA, 2001. Ten Years of Protein Kinase B Signalling: A Hard Akt to Follow. Trends in Biochemical Sciences, 26(11): 657-664. doi: 10.1016/s0968-0004(01)01958-2.
  • Dede S, Yur F, Taşpınar M, Çetin S, Usta A, Yüksek V, 2019. In Vitro Evaluation of Thymoquinone and Lycopene Supplementation on Oxidative DNA Damage and Oxidant Status in High Glucose Conditions. Latin American Journal Pharmacy, 38 (1): 209-912.
  • Ghosheh OA, Houdi A, Crooks PA, 1999. High Performance Liquid Chromatography Analysis of The Pharmacologically Active Quinines and Related Compounds in The Oil of The Black Seed (Nigella Sativa). Journal of Pharmaceutical and Biomedical Analysis, 19(5): 757-762.
  • Hemmings BA, Restuccia DF, 2015. PI3K-PKB/Akt Pathway. Cold Spring Harbor Perspectives in Biology, 1;4(9): a011189. doi: 10.1101/cshperspect.a011189.
  • Hosseinzadeh H, Taiari S, Nassiri-Asl M, 2012. Effect of Thymoquinone, a Constituent of Nigella Sativa L., On İschemia–Reperfusion İn Rat Skeletal Muscle. Naunyn-Schmiedeberg's Archives of Pharmacology, 385(5), 503-508.
  • Iskender B, Izgi K, Canatan H, 2016. Novel Anti-Cancer Agent Myrtucommulone-A And Thymoquinone Abrogate Epithelial-Mesenchymal Transition in Cancer Cells Mainly Through The İnhibition of PI3K/AKT Signalling Axis. Molecular and Cellular Biochemistry, 416(1-2):71-84. doi: 10.1007/s11010-016-2697-y.
  • Khader M, Eckl PM, 2014. Thymoquinone: An Emerging Natural Drug withth a Wide Range of Medical Applications. Iran Journal Basic Medicine Science, 17(12): 950-957
  • Porta C, Paglino C, Mosca A, 2014. Targeting PI3K/Akt/mTOR Signaling in Cancer. Frontiers in Oncology, 14;4:64. doi: 10.3389/fonc.2014.00064.
  • Xu D, Ma Y, Zhao B, Li S, Zhang Y, Pan S, Wu Y, Wang J, Wang D, Pan H, Liu L, Jiang H, 2014. Thymoquinone Induces G2/M Arrest, Inactivates PI3K/Akt and Nuclear factor-κB Pathways in Human Cholangiocarcinomas both in vitro and in vivo. Oncology Reports, 31(5):2063-2070. doi: 10.3892/or.2014.3059..
  • Yang XL, Lin FJ, Guo YJ, Shao ZM, Ou ZL, 2014. Gemcitabine Resistance in Breast Cancer Cells Regulated By PI3K/AKT-Mediated Cellular Proliferation Exerts Negative Feedback via The MEK/MAPK and mTOR Pathways. OncoTargets and Therapy, 7:1033-1042. doi: 10.2147/OTT.S63145.
  • Yüksek V, Dede S, Taşpınar M, Yur F, Çetin S, 2017. Effects of Thymoquınone Applicatıon in The Renal Cell Line. International Congress on Medicinal and Aromatic Plants, Konya, May 10-12, 2017, pp: 842-844.
There are 13 citations in total.

Details

Primary Language Turkish
Subjects Structural Biology
Journal Section Biyoloji / Biology
Authors

Veysel Yüksek 0000-0001-7432-4989

Publication Date March 1, 2021
Submission Date October 28, 2020
Acceptance Date November 12, 2020
Published in Issue Year 2021 Volume: 11 Issue: 1

Cite

APA Yüksek, V. (2021). NRK-52E Hücre Serisinde Timokinon İndüklü PI3K/AKT/mTOR Yolak Aktivasyonu. Journal of the Institute of Science and Technology, 11(1), 68-74. https://doi.org/10.21597/jist.817666
AMA Yüksek V. NRK-52E Hücre Serisinde Timokinon İndüklü PI3K/AKT/mTOR Yolak Aktivasyonu. J. Inst. Sci. and Tech. March 2021;11(1):68-74. doi:10.21597/jist.817666
Chicago Yüksek, Veysel. “NRK-52E Hücre Serisinde Timokinon İndüklü PI3K/AKT/MTOR Yolak Aktivasyonu”. Journal of the Institute of Science and Technology 11, no. 1 (March 2021): 68-74. https://doi.org/10.21597/jist.817666.
EndNote Yüksek V (March 1, 2021) NRK-52E Hücre Serisinde Timokinon İndüklü PI3K/AKT/mTOR Yolak Aktivasyonu. Journal of the Institute of Science and Technology 11 1 68–74.
IEEE V. Yüksek, “NRK-52E Hücre Serisinde Timokinon İndüklü PI3K/AKT/mTOR Yolak Aktivasyonu”, J. Inst. Sci. and Tech., vol. 11, no. 1, pp. 68–74, 2021, doi: 10.21597/jist.817666.
ISNAD Yüksek, Veysel. “NRK-52E Hücre Serisinde Timokinon İndüklü PI3K/AKT/MTOR Yolak Aktivasyonu”. Journal of the Institute of Science and Technology 11/1 (March 2021), 68-74. https://doi.org/10.21597/jist.817666.
JAMA Yüksek V. NRK-52E Hücre Serisinde Timokinon İndüklü PI3K/AKT/mTOR Yolak Aktivasyonu. J. Inst. Sci. and Tech. 2021;11:68–74.
MLA Yüksek, Veysel. “NRK-52E Hücre Serisinde Timokinon İndüklü PI3K/AKT/MTOR Yolak Aktivasyonu”. Journal of the Institute of Science and Technology, vol. 11, no. 1, 2021, pp. 68-74, doi:10.21597/jist.817666.
Vancouver Yüksek V. NRK-52E Hücre Serisinde Timokinon İndüklü PI3K/AKT/mTOR Yolak Aktivasyonu. J. Inst. Sci. and Tech. 2021;11(1):68-74.