Exosomes from Von Hippel-Lindau-Null Cancer Cells Promote Metastasis in Renal Cell Carcinoma
Abstract
:1. Introduction
2. Results
2.1. Physical and Biochemical Characterization of RCC Exosomes
2.2. VHL(−) Exosomes Promote VHL(+) Migration and Invasion
2.3. Exosome Uptake
2.4. Visualization of Exosome Internalization and Induced Migration Using a Cre-loxP System
2.5. VHL(−) Exosomes Promote RCC Metastasis in Ovo
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Exosome Isolation, Imaging, and Quantification
4.3. Internalization of PKH67-Labeled VHL-Knockout Exosomes
4.4. Cre-loxP Color Conversion Assay
4.5. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
RCC | Renal cell carcinoma |
ccRCC | clear cell renal cell carcinoma |
CAM | Chorioallantoic membrane model |
DLS | Dynamic light scattering |
DAPI | 4′,6-diamidino-2-phenylindole |
GAPDH | Glyceraldehyde-3-phosphate dehydrogenase |
BCA | Bicinchoninic acid |
qPCR | quantitative PCR |
RT-qPCR | Reverse transcription qPCR |
SD | Standard deviation |
MMP-9 | Matrix metalloproteinase 9 |
EMT | Epithelial–mesenchymal transition |
RFP | Red fluorescent protein |
GFP | Green fluorescent protein |
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Cell Line | Parental Cells | Modifications |
---|---|---|
RC-VHL(+) | Renca | VHL wildtype, RFP; as described in [15] |
RC-VHL(−) | Renca | VHL knockout, GRP; as described in [15] |
AC-VHL(+) | ACHN | VHL wildtype; as described in [15] |
AC-VHL(−) | ACHN | VHL knockout; as described in [15] |
RC-VHL(+)-loxP | RC-VHL(+) | RFP → GFP upon receiving Cre; lentivirally overexpressed plasmid from Addgene#65726 [33,34] |
RC-VHL(−)-Cre | RC-VHL(−) | Lentivirally overexpressed plasmid from Addgene#65727 [33,34] |
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Flora, K.; Ishihara, M.; Zhang, Z.; Bowen, E.S.; Wu, A.; Ayoub, T.; Huang, J.; Cano-Ruiz, C.; Jackson, M.; Reghu, K.; et al. Exosomes from Von Hippel-Lindau-Null Cancer Cells Promote Metastasis in Renal Cell Carcinoma. Int. J. Mol. Sci. 2023, 24, 17307. https://doi.org/10.3390/ijms242417307
Flora K, Ishihara M, Zhang Z, Bowen ES, Wu A, Ayoub T, Huang J, Cano-Ruiz C, Jackson M, Reghu K, et al. Exosomes from Von Hippel-Lindau-Null Cancer Cells Promote Metastasis in Renal Cell Carcinoma. International Journal of Molecular Sciences. 2023; 24(24):17307. https://doi.org/10.3390/ijms242417307
Chicago/Turabian StyleFlora, Kailey, Moe Ishihara, Zhicheng Zhang, Elizabeth S. Bowen, Aimee Wu, Tala Ayoub, Julian Huang, Celine Cano-Ruiz, Maia Jackson, Kaveeya Reghu, and et al. 2023. "Exosomes from Von Hippel-Lindau-Null Cancer Cells Promote Metastasis in Renal Cell Carcinoma" International Journal of Molecular Sciences 24, no. 24: 17307. https://doi.org/10.3390/ijms242417307