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Published on: March 30, 2019
[miR-520a-3p regulates the signaling pathway of cytokine secretion in cervical cancer]
Hong-Yan Liu1, Qi-Hong Lu2, Chu-Ying Huang3
1Tumor Radiotherapy Center of Central Hospital of Enshi Tujia Miao Autonomous Prefecture, Hubei Province, Enshi 445000.
Objective:
To investigate the molecular mechanism of miR-520a-3p regulating the secretion of cytokines in cervical cancer.
Methods:
The matching between miR-520a-3p and the NF-κB complex subunit RELA was analyzed by TargetScanHuman, and then the luciferase reporting system was used to detect whether miR-520a-3p targeted the NF-κB complex subunit RELA. After cervical cancer HELA cells stimulated by LPS, in the overexpression group or the knockout group, miR-520a-3p mimics or inhibitor were mixed with transfection reagent and dripped into HELA cells. The expression levels of colony stimulating factor (GCSF), granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin 2 (IL-2), interleukin 3 (IL-3), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 19 (IL-9), interleukin 10 (IL-10), interleukin 12 p40 (IL-12 p40), interleukin 12 p70 (IL-12 p70), interleukin 13 (IL-13), interleukin 17 (IL-17), interferon gamma (IFN-γ), monocyte chemoattractant protein-1 (MCP-1), monocyte chemoattractant protein-5 (MCP-5), small inducible cytokine A5 (RANTES), tumor necrosis factor-alpha (TNFα) were detected by enzyme-linked immunosorbent assay in both the overexpression group and the knockout group. Each experiment was repeated three times.
Results:
miR-520a-3p targeted the 3'UTR of RELA. After activation of the NF-κB signaling pathway by lipopolysaccharide (LPS), the protein expression levels of cytokines GCSF, GM-CSF, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40, IL-12 p70, IL-13, IL-17, IFN-γ, MCP-1, MCP-5, RANTES, TNFα secreted by HELA cells of cervical cancer were increased significantly (P＜0.05). In the overexpression group, the protein expression level of RELA of NF-κB complex subunit was decreased, and the protein expression level of the above-mentioned cytokines secreted by HELA cells of cervical cancer were decreased (P＜0.05). In the knockdown group, the protein expression level of RELA, a subunit of NF-κB complex, and the protein expression level of the above-mentioned cytokines secreted by HELA cells of cervical cancer were increased (P＜0.05).
Conclusion:
miR-520a-3p inhibits cytokine secretion in HELA cells of cervical cancer by targeting RELA, a key molecule in the NF-κB signaling pathway.
Insights
MicroRNA-520a-3p inhibits cytokine secretion in cervical cancer cells by targeting RELA, a key molecule in the NF-κB pathway. This finding offers insights into the molecular mechanisms of cervical cancer progression and potential therapeutic strategies.
Area of Science:
- Molecular Biology
- Oncology
- Immunology
Background:
- Cervical cancer is a significant global health concern.
- Cytokine secretion plays a crucial role in cancer progression and immune response.
- MicroRNAs (miRNAs) are emerging as critical regulators in various cancers, including cervical cancer.
Purpose of the Study:
- To elucidate the molecular mechanism by which miR-520a-3p regulates cytokine secretion in cervical cancer cells.
- To investigate the direct interaction between miR-520a-3p and the NF-κB signaling pathway.
Main Methods:
- Bioinformatic analysis (TargetScanHuman) predicted the interaction between miR-520a-3p and RELA.
- Luciferase reporter assays confirmed the targeting of RELA by miR-520a-3p.
- HELA cervical cancer cells were stimulated with LPS, and miR-520a-3p mimics or inhibitors were used to modulate its expression, followed by ELISA detection of various cytokines and RELA protein levels.
Main Results:
- miR-520a-3p was confirmed to target the 3'UTR of RELA, a subunit of the NF-κB complex.
- LPS stimulation significantly increased the secretion of multiple cytokines (GCSF, GM-CSF, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40, IL-12 p70, IL-13, IL-17, IFN-γ, MCP-1, MCP-5, RANTES, TNFα).
- Overexpression of miR-520a-3p decreased RELA and cytokine levels, while knockdown of miR-520a-3p increased them, indicating an inhibitory role of miR-520a-3p.
Conclusions:
- miR-520a-3p functions as an inhibitor of cytokine secretion in cervical cancer HELA cells.
- This inhibition is mediated through the targeting of RELA, a critical component of the NF-κB signaling pathway.
- The findings highlight miR-520a-3p as a potential therapeutic target for managing cervical cancer.
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