Expression levels of the microRNA maturing microprocessor complex components; Drosha, Dicer, and DGCR8 in PBMCs from
Zeinab Tabrizi1,2, Reza Mansouri2, Saeed Aslani1
1Rheumatology Research Center, Tehran University of Medical Sciences, Tehran, Iran.
Objective/Aim:
Two major enzymes in the microRNA maturation process, Dicer and Drosha, as well as DGCR8, the assistant of Drosha, function in the microprocessor complex. In this survey, the mRNA expression profiles of Drosha, Dicer, and DGCR8 in peripheral blood mononuclear cells (PBMCs) from ankylosing spondylitis (AS) patients and healthy controls were measured.
Methods:
Forty patients with AS and 40 age and gender matched healthy individuals were included in the study. PBMCs were separated, total RNA content of the cells was isolated, and first strand cDNA was synthesized. Quantitative analysis was performed through real-time PCR using the SYBR Green gene expression master mix.
Results:
AS cases expressed the Drosha mRNA almost equal to that of healthy controls (Fold Change= -0.94; P= 0.200). However, both Dicer and DGCR8 mRNA expressions were downregulated in patients relative to healthy subjects (Fold Change= -0.54 and -0.60; P= 0.002 and 0.004, respectively).
Conclusion:
Our results suggest that downregulation of miRNA maturation components, namely Dicer and DGCR8 may be contributing in the pathogenesis procedure of AS.
Insights
MicroRNA maturation enzymes Dicer and DGCR8 mRNA are downregulated in ankylosing spondylitis (AS) patients. This suggests their reduced function may contribute to AS development.
Area of Science:
- Molecular Biology
- Immunology
- Genetics
Background:
- MicroRNA (miRNA) maturation involves key enzymes like Dicer and Drosha, along with DGCR8, forming the microprocessor complex.
- Ankylosing spondylitis (AS) is a chronic inflammatory disease with complex pathogenesis.
- Understanding the role of miRNA biogenesis in AS may reveal new therapeutic targets.
Purpose of the Study:
- To investigate the mRNA expression levels of Drosha, Dicer, and DGCR8 in peripheral blood mononuclear cells (PBMCs) of AS patients compared to healthy controls.
- To determine if alterations in these miRNA processing components are associated with AS.
Main Methods:
- Quantitative real-time PCR was used to measure the mRNA expression of Drosha, Dicer, and DGCR8.
- Peripheral blood mononuclear cells (PBMCs) were isolated from 40 AS patients and 40 healthy controls.
- SYBR Green gene expression master mix was utilized for accurate quantification.
Main Results:
- Drosha mRNA expression showed no significant difference between AS patients and healthy controls (P=0.200).
- Dicer mRNA expression was significantly downregulated in AS patients compared to controls (Fold Change=-0.54, P=0.002).
- DGCR8 mRNA expression was also significantly downregulated in AS patients (Fold Change=-0.60, P=0.004).
Conclusions:
- The study indicates a downregulation of Dicer and DGCR8 mRNA in AS patients.
- These findings suggest that impaired miRNA maturation due to reduced Dicer and DGCR8 may play a role in the pathogenesis of ankylosing spondylitis.
- Further research is warranted to elucidate the precise mechanisms.
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