Time dependent gene expression analysis of vitamin D3 (1,25 (OH)2 D3) treated HL–60 cells by Q-RT-PCR


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Kanlı A., Gülkaç M. D. , Savlı H.

3rd International Eurasian Conference on Biological and Chemical Sciences (EurasianBioChem 2020), Ankara, Türkiye, 19 - 20 Mart 2020, ss.31

  • Basıldığı Şehir: Ankara
  • Basıldığı Ülke: Türkiye
  • Sayfa Sayıları: ss.31

Özet

Time dependent gene expression analysis of vitamin D3 (1,25 (OH)2 D3) treated HL–60 cells by Q-RT-PCR
Aylin Kanlı1, Mehmet Doğan Gülkaç1, Hakan Savlı2
1 Department of Medical Biology - Faculty of Medicine - University of Kocaeli- Turkey
2 Department of Medical Genetics - Faculty of Medicine - University of Kocaeli- Turkey
E-mail: aylin.kanli@kocaeli.edu.tr aylinkanli@hotmail.com
Abstract
Objective: The treatment of Acute myelogenous leukemia (AML) with differential effect of 1alpha,25-dihydroxyvitamin
D(3) (1,25(OH)2D3) has been one of the popular research topics. We aimed to analyze expression of several genes related to
cell cycle regulation and apoptosis in realization of 1,25(OH)2D3-induced differentiation of HL-60 cells.
Methods: We analyzed 13 genes (TNFR1, Bcl-w, Bax, Bak, Caspase–6, Caspase–8, AIF, Survivin, Cdk1 (Cdc2), Cdk2,
Cdk4, Cyclin-D1 and Cyclin-E) for changes in expression associated with the apoptosis of human promyelocytic leukemia
HL-60 cells induced by 1,25(OH)2D3 at 18th, 36th, 48th and 72nd h, using quantitative real-time PCR.
Results: We did not significantly find down or up-regulated expression profiles at mentioned time points. In our
experiments, we observed decreased expression of TNFR1, Cdk-4, Cyclin-D1, Cyclin-E and Survivin genes at 72nd hours,
and increased expression of Caspase-8 and Bak genes. The tendency of cell cycle related genes to decrease from 18th to 72nd
hours suggests that the cells entered the process of differentiation from the proliferation process. The anti-apoptotic genes
Bcl-w and Survivin gene expressions generally follow a decreasing line, and the tendency to increase expression of Bak
gene and Caspase-8 suggest that the apoptotic process starts around 72nd hours.
Conclusion: In conclusion, our results suggest that the leukemic cells are exposed to the effect of vitamin D3 as expected,
the repetitive clonal development process ends, the differentiation is completed, and the apoptotic process has just begun.
Keywords: HL–60 cells, 1,25 (OH)2 D3, Cell Cycle, Differentiation, Apoptosis, Quantitative Real Time Polymerase Chain
Reaction (Q-RT-PCR)
Acknowledgement: This research was supported by the Scientific Research Foundation of Kocaeli University. (Project
Number: 2004/31). This study is from a PhD thesis carried out at Kocaeli University by Aylin KANLI and her mentors
Professor Mehmet Doğan GÜLKAÇ and Professor Hakan SAVLI.