Related Experiment Video
Updated: Jul 12, 2026
Three-Dimensional Bone Extracellular Matrix Model for Osteosarcoma
Published on: April 12, 2019
The miR-16-1-3p passenger strand exhibits functional activity and suppresses malignant phenotypes in osteosarcoma
Wenyu Xue1, Yuzhe Wang1, Polina Pugacheva2
1Institute of Future Biophysics, Moscow Center for Advanced Studies, Moscow, Russia.
Background:
Chemotherapy resistance and disease progression remain major causes of mortality in osteosarcoma (OS). The miR-16 family is linked to tumors, but the specific function and treatment potential of the miR-16-1-3p passenger strand in OS remain unclear.
Methods:
RNA sequencing and clinical data from 82 OS patients in the TARGET-OS cohort were analyzed. Patients were stratified by progression status. Target genes of miR-16-1-3p were predicted using TargetScan and evaluated by gene set enrichment analysis (GSEA). Principal component analysis (PCA) and Kaplan-Meier survival analyses were performed based on cumulative Z-score-derived expression of the target gene set. Functional enrichment was assessed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. The impact of lentivirus-transduced miR-16-1-3p on U2OS cells was examined, focusing on proliferation, cell cycle, migration, colony formation, cisplatin sensitivity, and tumor growth in the CAM in vivo model. A fluorescent miRNA sensor was developed to assess intracellular target-binding functionality of the miR-16-1-3p passenger strand. Direct regulation of candidate target gene expression was validated using luciferase reporter assays and quantitative PCR. Expression of validated targets was further examined in a limited cohort of 15 chemotherapy-treated OS patients.
Results:
GSEA demonstrated significant enrichment of predicted miR-16-1-3p target genes among genes upregulated in patients with progressive disease. Kaplan-Meier analysis showed that elevated cumulative expression of predicted miR-16-1-3p target genes was associated with poorer overall survival in osteosarcoma patients. GO and KEGG analyses revealed enrichment of pathways related to translational regulation, mitochondrial function, and chemotherapy-associated signaling. A fluorescent miR-16-1-3p sensor system demonstrated sequence-dependent suppression of reporter activity following miR-16-1-3p overexpression. In functional assays performed in both U2OS and HOS cells, miR-16-1-3p overexpression significantly suppressed proliferation and migration and increased cisplatin sensitivity compared with control (scrambled miRNA) virus-transduced cells. Quantitative PCR and luciferase reporter assays further supported the interaction of miR-16-1-3p with SLC38A1 and ABCA13, two candidate genes potentially associated with chemoresistance.
Conclusion:
The miR-16-1-3p passenger strand exhibited functional activity associated with tumor-suppressive phenotypes and increased cisplatin sensitivity in osteosarcoma cells. Using integrative bioinformatics, fluorescent sensor assays, and functional experiments in U2OS and HOS cells points to miR-16-1-3p possibly regulating cancer characteristics and how osteosarcoma reacts to chemotherapy.
Insights
The miR-16-1-3p passenger strand shows tumor-suppressive activity in osteosarcoma (OS), enhancing cisplatin sensitivity. This microRNA may offer new therapeutic strategies for OS patients resistant to chemotherapy.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- Chemotherapy resistance and disease progression are critical challenges in osteosarcoma (OS) treatment.
- The role of the miR-16 family in OS is recognized, but the specific function of the miR-16-1-3p passenger strand remains largely unknown.
- Understanding miR-16-1-3p's function is crucial for developing novel therapeutic approaches in OS.
Purpose of the Study:
- To investigate the functional role and therapeutic potential of the miR-16-1-3p passenger strand in osteosarcoma.
- To identify target genes regulated by miR-16-1-3p and their association with OS progression and survival.
- To evaluate the impact of miR-16-1-3p on OS cell behavior and chemosensitivity.
Main Methods:
- Analysis of RNA sequencing and clinical data from 82 OS patients (TARGET-OS cohort).
- Bioinformatic prediction of miR-16-1-3p targets, pathway analysis (GSEA, GO, KEGG), and survival analysis (Kaplan-Meier).
- In vitro functional assays (proliferation, migration, cell cycle, cisplatin sensitivity) and in vivo CAM model using miR-16-1-3p overexpressed U2OS cells; validation of target interactions via luciferase reporter assays and qPCR.
Main Results:
- Enriched miR-16-1-3p target genes were upregulated in progressive OS, correlating with poorer survival.
- miR-16-1-3p overexpression suppressed proliferation and migration while increasing cisplatin sensitivity in OS cells.
- miR-16-1-3p demonstrated sequence-dependent target suppression, with SLC38A1 and ABCA13 identified as potential direct targets.
Conclusions:
- The miR-16-1-3p passenger strand exhibits tumor-suppressive properties and enhances cisplatin sensitivity in osteosarcoma.
- miR-16-1-3p plays a role in regulating osteosarcoma cell characteristics and response to chemotherapy.
- These findings suggest miR-16-1-3p as a potential therapeutic target for improving osteosarcoma treatment outcomes.
Related Concept Videos
02:23Abnormal Proliferation
03:03mTOR Signaling and Cancer Progression
The mTOR pathway or the...
01:22MicroRNAs
01:12Loss of Tumor Suppressor Gene Functions
When the tumor suppressor genes develop mutations or are lost, cells start growing out of control, leading to cancer. However, a single functional copy of the tumor suppressor gene is enough for the cells to maintain their normal functions and cell...