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Published on: April 6, 2012
MicroRNA-183-5p negatively regulates interleukin-8 expression in cervical cancer cells
1Department of Pathology, College of Medicine, Qassim University, Buraidah, Saudi Arabia.
Objectives:
Interleukin-8 (IL-8) and microRNA-183-5p (hsa-miR-183-5p) have been implicated in the development of cervical cancer, yet their relationship has not been explored. This study aims to determine whether phorbol 12-myristate 13-acetate (PMA)-induced IL-8 expression is regulated by hsa-miR-183-5p in cervical cancer cells.
Methods:
Bioinformatics algorithms were employed to predict the potential binding of hsa-miR-183-5p to the 3'UTR of IL-8 mRNA. CaSKi cervical cancer cells were used as a model to investigate this regulation. The expression levels of hsa-miR-183-5p and IL-8 were measured using Taqman assays through real-time polymerase chain reaction, while IL-8 protein levels were quantified in culture media through IL-8 specific Sandwich enzyme-linked immunosorbent assays. Luciferase reporter assays and transfections with pre- or anti-miR-183-5p were conducted to validate the binding of hsa-miR-183-5p to IL-8 mRNA's 3'UTR.
Results:
The bioinformatics tool TargetScan identified a seed-matched sequence for hsa-miR-183-5p in the 3'UTR of IL-8 mRNA. PMA-induced IL-8 expression was inversely correlated with hsa-miR-183-5p down regulation in cervical cancer cells. hsa-miR-183-5p significantly reduced luciferase activity in the 3'UTR-IL-8 reporter assay. Transfection with pre-miR-183-5p led to a notable decrease in IL-8 mRNA and protein secretion, while anti-miR-183-5p transfection caused a significant increase in IL-8 mRNA and protein levels in PMA-treated cells.
Conclusion:
This study is the first to demonstrate that hsa-miR-183-5p directly regulates IL-8 expression in cervical cancer cells. Both IL-8 and hsa-miR-183-5p could serve as potential therapeutic targets in the treatment of cervical cancer.
Insights
MicroRNA-183-5p (hsa-miR-183-5p) directly regulates Interleukin-8 (IL-8) expression in cervical cancer cells. This finding suggests both hsa-miR-183-5p and IL-8 are potential therapeutic targets for cervical cancer treatment.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Interleukin-8 (IL-8) and microRNA-183-5p (hsa-miR-183-5p) are implicated in cervical cancer development.
- The regulatory relationship between IL-8 and hsa-miR-183-5p in this context remains unexplored.
Purpose of the Study:
- To investigate whether phorbol 12-myristate 13-acetate (PMA)-induced IL-8 expression is regulated by hsa-miR-183-5p in cervical cancer cells.
- To elucidate the direct interaction between hsa-miR-183-5p and IL-8 mRNA.
Main Methods:
- Bioinformatic prediction of hsa-miR-183-5p binding to the IL-8 mRNA 3'UTR.
- Quantification of hsa-miR-183-5p and IL-8 mRNA/protein levels in CaSKi cervical cancer cells using Taqman assays and ELISA.
- Validation through luciferase reporter assays and transfection with pre-miR-183-5p and anti-miR-183-5p.
Main Results:
- hsa-miR-183-5p was predicted to bind to the IL-8 mRNA 3'UTR.
- PMA-induced IL-8 expression showed an inverse correlation with hsa-miR-183-5p levels.
- hsa-miR-183-5p significantly inhibited IL-8 expression at both mRNA and protein levels, confirmed by reporter assays and transfection studies.
Conclusions:
- This study provides the first evidence that hsa-miR-183-5p directly regulates IL-8 expression in cervical cancer.
- Both IL-8 and hsa-miR-183-5p represent promising therapeutic targets for cervical cancer intervention.
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