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Evaluation of the Relationship between Aromatase/Sirtuin1 Interaction and miRNA Expression in Human Neuroblastoma
Yasemin Kartal1,2, Unal Metin Tokat3, Pelin Kelicen-Ugur4
1Faculty of Medicine, Department of Physiology, Hacettepe University, Ankara, Turkey.
Background:
Changes in activation/inhibition of Sirtuin-1 (SIRT1) and aromatase play an important role in a plethora of diseases. MicroRNAs (miRNAs) modulate multiple molecular pathways and affect a substantial number of physiological and pathological processes.
Objective:
The aim of this study was to investigate any possible interaction between aromatase and SIRT1 in SH-SY5Y cells and to see how there is a connection between this interaction and miRNA expression, if there is an interaction.
Methods:
In this study, cells were incubated in serum-deprived media for 6, 12, and 24 h. Aromatase and SIRT1 expressions were evaluated by Western blot. The IC50 concentration of SIRT1 activator (SRT1720), SIRT1 inhibitor (EX527), and aromatase inhibitors (letrozole and fadrozole) was determined by the XTT method. Then, CYP19A1 and SIRT1 levels were evaluated in the presence of SIRT1 siRNA or IC50 values for each activator/inhibitor. Finally, CYP19A1, SIRT1 expression and miRNA target gene were assessed with bioinformatic approaches.
Results:
Aromatase and SIRT1 protein levels were significantly elevated in the cells incubated at 24 h in serum-deprived media (p ≤ 0.05). SIRT1 also positively regulated CYP19A1 in SH-SY5Y cells in media with/without FBS. Serum deprivation depending on time course caused changes in the oxidant/ antioxidant system. While oxidative stress index tended to decrease in the absence of FBS at 24 h compared to the control, it showed a significant decrease at 48 h in a serum-deprived manner (p ≤ 0.001). As a result of bioinformatics analysis, we determined 3 miRNAs that could potentially regulate SIRT1 and CYP19A1. hsa-miR-27a-3p and hsa-miR-181a-5p correlated in terms of their expressions at 24 h compared to 12 h, and there was a significant decrease in the expression of these miRNAs. On the contrary, the expression of hsa-miR-30c-5p significantly increased at 24 h compared to 12 h.
Conclusion:
Considering the results, a direct link between aromatase and SIRT1 was observed in human neuroblastoma cells. The identification of key miRNAs, hsa-miR-27a-3p, hsa-miR-30c-5p, and hsa-miR-181a-5p targeting both aromatase and SIRT1, provides an approach with novel insights on neurology-associated diseases.
Insights
This study reveals a direct link between aromatase and Sirtuin-1 (SIRT1) in neuroblastoma cells. Three specific microRNAs (miRNAs) were identified as key regulators of both aromatase and SIRT1, offering new insights into neurological diseases.
Area of Science:
- Neuroscience and Molecular Biology
- Cellular and Molecular Medicine
Background:
- Sirtuin-1 (SIRT1) and aromatase are crucial in various diseases.
- MicroRNAs (miRNAs) significantly influence physiological and pathological processes by modulating molecular pathways.
Purpose of the Study:
- To investigate the interaction between aromatase and SIRT1 in SH-SY5Y cells.
- To explore the connection between this interaction and miRNA expression patterns.
Main Methods:
- SH-SY5Y cells were cultured in serum-deprived media for varying durations.
- Protein expression of aromatase and SIRT1 was assessed using Western blot.
- Inhibitors, activators, and siRNA were used to modulate SIRT1 and aromatase activity.
- Bioinformatic analyses identified potential miRNA regulators of SIRT1 and aromatase (CYP19A1).
Main Results:
- Aromatase and SIRT1 protein levels increased significantly after 24 hours of serum deprivation.
- SIRT1 positively regulated CYP19A1 expression in SH-SY5Y cells.
- Serum deprivation altered the oxidant/antioxidant system, with a significant decrease in oxidative stress index at 48 hours.
- Three miRNAs (hsa-miR-27a-3p, hsa-miR-181a-5p, hsa-miR-30c-5p) were identified as potential regulators of SIRT1 and CYP19A1, with distinct expression changes observed.
Conclusions:
- A direct interaction between aromatase and SIRT1 was confirmed in human neuroblastoma cells.
- The identified miRNAs (hsa-miR-27a-3p, hsa-miR-30c-5p, hsa-miR-181a-5p) target both aromatase and SIRT1.
- These findings offer novel perspectives for understanding and potentially treating neurology-associated diseases.
