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Updated: Dec 27, 2025
Clinicopathological Analysis of miRNA Expression in Breast Cancer Tissues by Using miRNA In Situ Hybridization
Published on: June 7, 2016
MiRNA-96-5p impacts the progression of breast cancer through targeting FOXO3
Ziyi Yin1, Wenyan Wang1, Gengbao Qu1
1Department of Breast Surgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China.
Background:
Breast cancer is the most common malignant tumor in women worldwide, with a high mortality rate. MicroRNAs are small non-coding RNAs that negatively regulate the expression of target genes by interacting with the target gene 3'-UTR, and participate in cell differentiation, proliferation, apoptosis and metabolism. The function of miRNA-96-5p in the progression of breast cancer has not been reported.
Methods:
We used the StarBase database to investigate the expression of miRNA-96-5p in breast cancer and adjacent normal tissues. FOXO3 3'-UTR construct and luciferase reporter assays was performed for the target gene. Expression levels of miRNAs including its target were analyzed by qRT-PCR and western blot. Cell proliferation was detected by CCK8 and colony formation, EdU assay.
Results:
Luciferase reporter assays showed miRNA-96-5p directly targeted FOXO3. Abrogation of miRNA-96-5p by transfection with its inhibitors in breast cancer cells significantly suppressed miRNA-96-5p expression and breast cancer cells proliferation. Western blot revealed that overexpression of miRNA-96-5p substantially reduced FOXO3 protein expression. We used the GEPIA, UALCAN and KM-plotter databases to investigate the expression of FOXO3 in human breast cancer and adjacent normal tissues, and its correlation with survival. In addition, we found that FOXO3 spoiled miR-96-5p induced breast cancer cell proliferation block effecting.
Conclusions:
miRNA-96-5p may exert a tumor promotion role through negatively regulating tumor suppressor gene FOXO3 and promoting cell proliferation.
Insights
MicroRNA-96-5p promotes breast cancer progression by targeting the tumor suppressor FOXO3. Inhibiting this microRNA (miRNA) suppressed cancer cell proliferation, indicating a potential therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Breast cancer is a leading cause of cancer mortality in women globally.
- MicroRNAs (miRNAs) are key regulators of gene expression involved in cellular processes.
- The role of miRNA-96-5p in breast cancer progression was previously uncharacterized.
Purpose of the Study:
- To investigate the function of miRNA-96-5p in breast cancer.
- To identify the downstream targets of miRNA-96-5p in breast cancer cells.
- To explore the therapeutic potential of targeting miRNA-96-5p.
Main Methods:
- Utilized the StarBase database for miRNA expression analysis.
- Performed luciferase reporter assays to validate miRNA-target interactions.
- Assessed cell proliferation using CCK8, colony formation, and EdU assays.
- Analyzed protein expression via Western blot and gene expression using qRT-PCR.
Main Results:
- miRNA-96-5p directly targets and downregulates the tumor suppressor FOXO3.
- Inhibition of miRNA-96-5p significantly reduced breast cancer cell proliferation.
- FOXO3 expression was inversely correlated with miRNA-96-5p levels.
- FOXO3 overexpression counteracted the inhibitory effects of miRNA-96-5p on cell proliferation.
Conclusions:
- miRNA-96-5p acts as an oncomiR in breast cancer.
- The miRNA-96-5p/FOXO3 axis plays a crucial role in breast cancer cell proliferation.
- Targeting miRNA-96-5p may offer a novel therapeutic strategy for breast cancer treatment.
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