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Updated: Sep 27, 2025
Intramucosal Inoculation of Squamous Cell Carcinoma Cells in Mice for Tumor Immune Profiling and Treatment Response Assessment
Published on: April 22, 2019
miR-211-3p enhances induction chemotherapy insensitivity by upregulating CSF2/CCL20/TNF signaling in hypopharyngeal
Lingwa Wang1, Ru Wang1, Tianqiao Huang2
1Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing, 100730, China.
Purpose:
To investigate the potential mechanisms of miR-211-3p on induction chemotherapy (IC) sensitivity in hypopharyngeal squamous cell carcinoma (HSCC).
Methods:
qRT-PCR was assessed to compare the miR-211-3p expression between IC sensitive and insensitive tumor tissues. The MTT assay was performed to analyze cell proliferation and viability to paclitaxel after alteration of miR-211-3p. Flow cytometry assay was conducted to explore cell apoptosis. Transwell assay was used to explore the effect of miR-211-3p on cell migration. Transcriptome sequencing was then performed to select differentially expressed genes (DEGs) after over-expression of miR-211-3p. GO and KEGG enrichment analyses were conducted to annotate DEGs. PPI analysis was conducted to screen candidate genes. The differential expression and survival status of candidate genes were further validated in TCGA-HNSCC data. The single sample GSEA method was used to investigate the association between downstream genes and immune cell infiltration.
Results:
miR-211-3p was up-regulated in IC insensitive larynx-hypopharyngeal tumor tissues. Over-expression of miR-211-3p promoted cell proliferation and migration, and inhibited apoptosis. The IC50 value of miR-211-3p overexpression (OE) group was significantly higher than negative control (NC) group treated with paclitaxel, suggesting miR-211-3p enhanced IC insensitivity in HSCC. We found 778 DEG after over-expression of miR-211-3p and 11 significant genes were then identified. Finally, colony stimulating factor 2 (CSF2) and C-C motif chemokine ligand 20 (CCL20) were validated to be significantly high expressed and associated with poorer overall survival in head and neck squamous cell carcinoma, which were involved in TNF signaling pathway and then regulated immune cell infiltration.
Conclusion:
The miR-211-3p could promote HSCC progression and upregulate CSF2/CCL20/TNF signaling to promote IC insensitivity in HSCC, which may provide new ideas for HSCC therapy.
Insights
MicroRNA-211-3p promotes hypopharyngeal squamous cell carcinoma progression and resistance to chemotherapy. Upregulation of miR-211-3p enhances tumor growth and immune evasion, suggesting it as a therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Hypopharyngeal squamous cell carcinoma (HSCC) poses significant treatment challenges.
- Understanding molecular mechanisms of chemoresistance is crucial for improving patient outcomes.
Purpose of the Study:
- To elucidate the role of microRNA-211-3p (miR-211-3p) in chemoresistance of HSCC.
- To investigate the downstream signaling pathways influenced by miR-211-3p.
Main Methods:
- Quantitative reverse transcription PCR (qRT-PCR) for miR-211-3p expression analysis.
- Cell proliferation, apoptosis, and migration assays (MTT, flow cytometry, Transwell).
- Transcriptome sequencing, Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and protein-protein interaction (PPI) analyses.
Main Results:
- miR-211-3p was upregulated in chemoinsensitive HSCC tissues.
- Overexpression of miR-211-3p increased proliferation, migration, and chemoresistance, while inhibiting apoptosis.
- CSF2 and CCL20 were identified as key downstream genes involved in the TNF signaling pathway, correlating with poor survival and immune cell infiltration.
Conclusions:
- miR-211-3p promotes HSCC progression and chemoresistance by upregulating the CSF2/CCL20/TNF signaling pathway.
- Targeting miR-211-3p or its downstream effectors may offer novel therapeutic strategies for HSCC.
