Related Experiment Video
Updated: Sep 25, 2025
Preparation of Exosomes for siRNA Delivery to Cancer Cells
Published on: December 5, 2018
In vitro experimental study on the formation of microRNA-34a loaded exosomes and their inhibitory effect in oral
Wei Deng1,2, Ying Meng1,2, Bin Wang1,3
1Jiangsu Key Laboratory of Oral Disease,Nanjing Medical University, Nanjing, Jiangsu, China.
Abstract:
Studies have shown the inhibitory effect of microRNA-34a on proliferation, migration, and invasion of oral squamous cell carcinoma. However, the lack of a safe and effective delivery system limits the clinical application of microRNA-34a in oral cancer treatment. An exosome is a small extracellular vesicle that mediates intercellular communication by delivering proteins, nucleic acids, and other contents, and functions as a natural drug delivery carrier. Here, we aimed to explore whether exosomes could be used to load microRNA-34a via co-incubation and further used to treat OSCC. Ultracentrifugation was used to obtain exosomes derived from HEK293T cells and the extracted exosomes were analyzed via transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blotting. Subsequently, we loaded cholesterol-modified microRNA-34a into HEK293T cell exosomes by co-incubation. Then, PKH67 and Cy3 co-labeled exo-microRNA-34a were co-incubated with HN6 cells and exosome entry into the HN6 cells was observed using a confocal laser scanning microscope. The cell proliferation, migration, and invasion were assessed by CCK-8 and Transwell assay analysis. SATB2 expression in HN6 cells was analyzed via western blotting. In this study, cholesterol-modified microRNA-34a was loaded into exosomes of HEK293T cells by co-incubation. The microRNA-34a-loaded exosomes were secreted from HEK293T cells and were absorbed by HN6 oral squamous carcinoma cells. Further, microRNA-34a-loaded exosomes led to a significant inhibition of HN6 cell proliferation, migration, and invasion by down regulating SATB2 expression. These results report a new delivery method for microRNA-34a, providing a new approach for the treatment of oral cancer.
Insights
Researchers developed a novel method to deliver microRNA-34a using exosomes for oral cancer treatment. This exosome-based delivery effectively inhibited oral squamous cell carcinoma proliferation, migration, and invasion.
Area of Science:
- Biomedical Engineering
- Molecular Oncology
- Nanotechnology
Background:
- MicroRNA-34a exhibits inhibitory effects on oral squamous cell carcinoma (OSCC) progression.
- Clinical application of microRNA-34a is hindered by the absence of effective delivery systems.
- Exosomes, as natural nanovesicles, offer potential as drug delivery vehicles for cancer therapy.
Purpose of the Study:
- To investigate the feasibility of using exosomes to deliver microRNA-34a for OSCC treatment.
- To develop a method for loading microRNA-34a into exosomes derived from HEK293T cells.
- To evaluate the therapeutic efficacy of exosome-mediated microRNA-34a delivery on OSCC cells.
Main Methods:
- Exosomes were isolated from HEK293T cells using ultracentrifugation and characterized by TEM, NTA, and Western blotting.
- Cholesterol-modified microRNA-34a was loaded into exosomes via co-incubation.
- Exosome uptake by HN6 OSCC cells was confirmed using confocal microscopy.
- Cell proliferation, migration, and invasion assays (CCK-8, Transwell) were performed.
- SATB2 expression was analyzed via Western blotting.
Main Results:
- Cholesterol-modified microRNA-34a was successfully loaded into HEK293T cell-derived exosomes.
- Exosomes efficiently delivered microRNA-34a into HN6 oral squamous carcinoma cells.
- MicroRNA-34a-loaded exosomes significantly inhibited HN6 cell proliferation, migration, and invasion.
- Downregulation of SATB2 expression was observed in HN6 cells treated with microRNA-34a-loaded exosomes.
Conclusions:
- Exosomes provide a viable and effective delivery system for microRNA-34a in oral cancer.
- This exosome-based approach demonstrates therapeutic potential for OSCC treatment by inhibiting tumor progression.
- The findings present a novel strategy for microRNA-34a delivery, advancing oral cancer therapy options.
Related Concept Videos
01:22MicroRNAs
01:36Overview of Exosomes
Stahl et al. discovered exosomes in 1983, but the exosomes were initially considered waste products released from the...