Exosomal Thrombospondin-1 Disrupts the Integrity of Endothelial Intercellular Junctions to Facilitate Breast Cancer Cell Metastasis
Abstract
:1. Introduction
2. Results
2.1. Exosome-Containing Fraction Derived from Cancer Cells Enhances the Transendothelial Migration of Breast Cancer Cells
2.2. Exosomes Enhance the Transendothelial Migration of MDA-MB-231 Cells and Inhibit the Tube Formation in HUVECs
2.3. Exosomes Derived from MDA-MB-231 Cells Suppress the Expression of Intercellular Junction Proteins in HUVECs
2.4. Tumor-Derived Exosomal TSP1 Promotes the Transendothelial Migration of Breast Cancer Cells In Vitro
2.5. Exosomal TSP1 Derived from Breast Cancer Cells Is Responsible for the Suppression of Intercellular Junction Molecules
2.6. Exosomal TSP1 Promotes the Migration of Tumor Cells in Zebrafish
3. Discussion
4. Materials and Methods
4.1. Cell Lines and LSKL Peptide Treatment
4.2. Preparation of Exosomes-Enriched Precipitate
4.3. Exosomes Preparation
4.4. Characterization of Exosomes
4.5. Western Blot
4.6. Immunofluorescence Analysis
4.7. Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) Analysis
4.8. Gene Cloning, Plasmid Construction and Transfection
4.9. Quantitative Real-Time PCR (qRT-PCR)
4.10. Transendothelial Migration Assay and Tube Formation
4.11. In Vivo Metastasis Assay
4.12. Bioinformatics Analysis
4.13. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Cen, J.; Feng, L.; Ke, H.; Bao, L.; Li, L.Z.; Tanaka, Y.; Weng, J.; Su, L. Exosomal Thrombospondin-1 Disrupts the Integrity of Endothelial Intercellular Junctions to Facilitate Breast Cancer Cell Metastasis. Cancers 2019, 11, 1946. https://doi.org/10.3390/cancers11121946
Cen J, Feng L, Ke H, Bao L, Li LZ, Tanaka Y, Weng J, Su L. Exosomal Thrombospondin-1 Disrupts the Integrity of Endothelial Intercellular Junctions to Facilitate Breast Cancer Cell Metastasis. Cancers. 2019; 11(12):1946. https://doi.org/10.3390/cancers11121946
Chicago/Turabian StyleCen, Junyu, Lingyun Feng, Huichuan Ke, Lifeng Bao, Lin Z. Li, Yoshimasa Tanaka, Jun Weng, and Li Su. 2019. "Exosomal Thrombospondin-1 Disrupts the Integrity of Endothelial Intercellular Junctions to Facilitate Breast Cancer Cell Metastasis" Cancers 11, no. 12: 1946. https://doi.org/10.3390/cancers11121946