Evaluation of anti-metastatic effect of chitosan nanoparticles on esophageal cancer-associated fibroblasts

Abstract

Aim: Esophageal cancer is one of the major types of cancers, causing death of approximately 5% of all cancer deaths. This is due, in large part, to both relatively ineffectual and unavailable treatment. In order to develop an effective treatment strategy against esophageal cancer, it is important to target metastatic genes. In the present study, we have used a cancer-associated fibroblast (CAF) cell line derived from culturing peripheral blood mononuclear cells from a metastatic esophageal cancer patient to see whether chitosan nanoparticles (Ch-Np) treatment can modulate the metastatic phenotype of CAF cells by using various cellular and molecular markers.

Methods: A CAF cell line was developed from peripheral blood mononuclear cells (PBMC) from a metastatic esophageal cancer patient. The cells were treated with 100 µg/mL of chitosan nanoparticle in vitro for the morphological and oncogenic characteristic studies, along with the expression of various genes involved in process of tumor development and metastasis. Techniques such as Light and Phase Contrast Microscopy, cell growth rate, Scratch metastatic assay, and molecular profiling were carried out to see changes in CAF cells before and after Ch-Np treatment.

Results: It was observed that CAF cells grew in monolayer and had a doubling time of 25 ± 0.38 h. Morphologically, the cells had a fibroblastic appearance. After treatment with 100 µg/mL of Ch-Np in vitro, there was an increased doubling time to 30 ± 0.83 h. Similarly, Scratch Assay showed an inhibition in the metastatic property of these cells. These findings were confirmed with gene expression studies. It was also observed that there was complete down-regulation of metastatic genes MMP1 and MMP9 and chemokines such as CXCR-4, CXCR-7, CCR-5, and SDF-1, indicating that Ch-Np inhibited the metastatic characteristic of CAF cells.

Conclusion: This study has shown that there was an inhibition of metastatic properties of CAF cells after treatment with Ch-Np, suggesting that Ch-Np can be a delivery system used for targeting cancer cells for treatment of esophageal cancer.

Keywords

Cancer-associated fibroblast / molecular markers / metastasis / chitosan nanoparticle / anti-metastatic / metastatic genes

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