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Tranilast induces MiR-200c expression through blockade of RelA/p65 activity in leiomyoma smooth muscle cells
Tsai-Der Chuang1, Amit Rehan1, Omid Khorram1
1Lundquist Institute for Biomedical Innovation and Department of Obstetrics and Gynecology, Harbor-University of California Los Angeles Medical Center, Torrance, California.
Objective:
To determine the mechanism by which tranilast induces miR-200c expression in leiomyoma smooth muscle cells (LSMCs).
Design:
Experimental study.
Setting:
Academic research laboratory.
Patient(S):
Women undergoing hysterectomy for leiomyoma.
Intervention(S):
Blockade of RelA/p65.
Main Outcome Measure(S):
Effects of tranilast and blockade of RelA/p65 on miR-200c expression.
Result(S):
Tranilast, an inflammation inhibitor, dose-dependently induced miR-200c in LSMCs and myometrium smooth muscle cells (MSMCs), with a more profound effect in LSMCs than in MSMCs. The treatment of LSMCs with Bay 117082, an inhibitor of IκB phosphorylation, further enhanced miR-200c induction by tranilast. The knockdown of RelA/p65 by small interfering RNA also induced miR-200c expression in LSMCs. Although tranilast had no effect on total RelA/p65 protein levels in LSMCs, it significantly induced RelA/p65 phosphorylation at S536 while reducing its activity as well as its nuclear translocation. ChIP assay indicated that tranilast reduces the binding ability of RelA/p65 to miR-200c promoter, resulting in miR-200c induction. Tranilast also inhibited interleukin-8 (IL8) expression in LSMCs. The induction of miR-200c by tranilast partially mediates the inhibitory effect of tranilast on the expression of IL8 and cyclin-dependent kinase 2 in LSMCs.
Conclusion(S):
Induction of miR-200c by tranilast in LSMCs is mediated through a transcriptional mechanism involving inhibition of the nuclear factor κB signaling pathway. These results highlight the significance of inflammation in the pathogenesis of leiomyoma and the potential utility of antiinflammatory drugs for treatment of leiomyomas.
Insights
Tranilast, an inflammation inhibitor, increases miR-200c in leiomyoma cells by blocking the NF-κB pathway. This finding suggests anti-inflammatory drugs may treat uterine fibroids.
Area of Science:
- Molecular biology
- Cell biology
- Pharmacology
Background:
- Uterine leiomyomas are common benign tumors.
- Inflammation plays a role in leiomyoma pathogenesis.
- MicroRNAs (miRNAs) are implicated in leiomyoma development.
Purpose of the Study:
- To elucidate the mechanism by which tranilast induces miR-200c expression in leiomyoma smooth muscle cells (LSMCs).
Main Methods:
- Experimental study using LSMCs and myometrium smooth muscle cells (MSMCs).
- Tranilast treatment and blockade of RelA/p65 (NF-κB pathway component) using Bay 117082 and small interfering RNA.
- Analysis of miR-200c expression, RelA/p65 phosphorylation, nuclear translocation, and promoter binding via ChIP assay.
- Measurement of IL8 and cyclin-dependent kinase 2 expression.
Main Results:
- Tranilast dose-dependently induced miR-200c in LSMCs and MSMCs, with a greater effect in LSMCs.
- Inhibition of IκB phosphorylation and RelA/p65 knockdown further enhanced miR-200c induction.
- Tranilast inhibited RelA/p65 activity and nuclear translocation, reducing its binding to the miR-200c promoter.
- Tranilast inhibited IL8 expression, and miR-200c induction partially mediated this effect and the inhibition of cyclin-dependent kinase 2.
Conclusions:
- Tranilast induces miR-200c in LSMCs via transcriptional inhibition of the NF-κB signaling pathway.
- These findings underscore the role of inflammation in leiomyoma and suggest anti-inflammatory drugs as potential therapeutics.
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