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Updated: May 29, 2025
Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
Mycophenolate mofetil reduces cell viability associated with the miR-205-PAX9 pathway in human lip fibroblast cells
Hiroki Yoshioka1,2, Hanane Horita1, Yosuke Tsukiboshi1
1Faculty of Pharmacy, Gifu University of Medical Science, 4-3-3 Nijigaoka, Kani, Gifu, 509-0293, Japan.
Abstract:
Cleft lip is a birth defect associated with environmental and genetic factors. Recently, microRNAs (miRNAs) have been reported to play a crucial role in lip formation, with the disruption of miRNAs influencing the development of cleft lip. Exposure to medicinal agents in pregnant women is one of the reasons for cleft lip. Although an association between pharmaceuticals-induced cleft lip and miRNAs has been suggested, it remains to be fully elucidated. This study aimed to clarify the molecular mechanism of mycophenolate mofetil (MPM)-induced inhibition of cell proliferation and miRNA expression in human lip fibroblast (KD) cells. Cell viability, apoptosis, and cell cycle-related markers were evaluated after 72 h of MPM treatment. In addition, miRNA levels and the expression of their downstream genes were measured, and a rescue experiment was performed by overexpressing PAX9. We showed that MPM dose-dependently reduced the viability of KD cells. In addition, MPM treatment suppressed cyclin-D1 and cyclin dependent kinase-6 expression in KD cells. Furthermore, MPM upregulated miR-205 expression and downregulated the expression of PAX9 (downstream gene). Moreover, PAX9 overexpression alleviated MPM-induced inhibition of cell proliferation. These results suggest that MPM suppresses cell viability by modulating miR-205-PAX9 expression.
Insights
Mycophenolate mofetil (MPM) reduces human lip fibroblast viability by upregulating miR-205 and downregulating PAX9. Overexpressing PAX9 reversed this MPM-induced inhibition, suggesting a novel molecular mechanism for drug-induced cleft lip.
Area of Science:
- Developmental Biology
- Molecular Biology
- Pharmacology
Background:
- Cleft lip is a birth defect influenced by genetic and environmental factors, including pharmaceutical exposure during pregnancy.
- MicroRNAs (miRNAs) are critical regulators of lip development, and their dysregulation is implicated in cleft lip.
- The precise molecular mechanisms linking pharmaceutical agents to miRNA alterations and cleft lip remain incompletely understood.
Purpose of the Study:
- To elucidate the molecular mechanism by which mycophenolate mofetil (MPM) inhibits cell proliferation and alters miRNA expression in human lip fibroblasts (KD cells).
- To investigate the role of miR-205 and its downstream target PAX9 in MPM-induced effects on cell viability.
Main Methods:
- Human lip fibroblast (KD) cells were treated with varying concentrations of MPM for 72 hours.
- Cell viability, apoptosis, and cell cycle markers were assessed.
- miRNA and downstream gene expression levels were quantified using quantitative real-time PCR.
- Rescue experiments involved overexpressing PAX9 to evaluate its impact on MPM-induced inhibition.
Main Results:
- MPM treatment dose-dependently decreased KD cell viability.
- MPM suppressed the expression of cell cycle regulators cyclin-D1 and cyclin dependent kinase-6.
- MPM upregulated miR-205 expression and downregulated its downstream target, PAX9.
- Overexpression of PAX9 attenuated the inhibitory effects of MPM on cell proliferation.
Conclusions:
- MPM suppresses cell viability in human lip fibroblasts by modulating the miR-205/PAX9 pathway.
- This study reveals a potential molecular mechanism for pharmaceutical-induced cleft lip, highlighting the role of specific miRNA-gene interactions.
