Comparative Study of Metastasis Suppression Effects of Extracellular Vesicles Derived from Anaplastic Cell Lines, Nanog-Overexpressing Melanoma, and Induced Pluripotent Stem Cells
Abstract
:1. Introduction
2. Results
2.1. Transfer of EVs to Various Organs
2.2. Effects of EVs as a Vaccine
2.3. Effects on Immune Cells
2.4. Results of miRNA Analysis
3. Discussion
4. Material and Methods
4.1. Cell Culture
4.2. Preparation of EVs
4.3. Western Analysis of CD81 and Gapdh
4.4. Analysis of the Transfer of EVs to Various Organs in a Mouse
4.5. In Vivo Test of the Effects of EVs on the Metastasis of Melanoma
4.6. Invasion Ability Test
4.7. Phagocytic Activity Test
4.8. Cytotoxic Activity Test
4.9. miRNA Analysis
4.10. Statistical Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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miRNA | Log2(FC) | Number of Target Genes | |||
---|---|---|---|---|---|
p < 0.05CWCS ≤ −0.4 | Targeted by Each miRNA among Top30 Genes | ||||
Nanog+F10-EVs/F10-EVs | miR-18a-5p | 2.46 | 42 | 301 | 4 |
miR-3473e | 1.75 | 6 | 0 | ||
miR-19a-3p | 1.20 | 126 | 11 | ||
miR-3473b | 1.18 | 127 | 5 | ||
miR-210-3p | −3.62 | 6 | 104 | 0 | |
miR-369-3p | −2.52 | 7 | 2 | ||
miR-122-5p | −1.79 | 29 | 5 | ||
miR-706 | −1.28 | 62 | 3 | ||
Sum | 405 | 30 | |||
iPS-EVs/Nanog+F10-EVs | miR-466h-3p | 7.62 | 58 | 275 | 1 |
miR-706 | 6.23 | 62 | 3 | ||
miR-323-3p | 6.23 | 10 | 0 | ||
miR-466f-3p | 6.07 | 145 | 5 | ||
miR-30a-3p | −4.99 | 49 | 394 | 0 | |
miR-342-5p | −4.74 | 335 | 20 | ||
let-7c-5p | −4.63 | 2 | 0 | ||
miR-423-3p | −4.36 | 8 | 1 | ||
Sum | 669 | 30 |
Gene | MCC Score | Degree | Regulation by mi RNA | |
---|---|---|---|---|
Nanog+F10-EVs | Trp53 | 1058 | 23 | up-regulated-miR-3473b |
Hif1a | 990 | 19 | up-regulated-miR-18a-5p | |
Esr1 | 919 | 11 | up-regulated-miR-18a-5p | |
Atm | 872 | 9 | up-regulated-miR-18a-5p | |
Cdkn1b | 847 | 7 | down-regulated-miR-706 | |
iPS-EVs | Ins1 | 13,801 | 19 | up-regulated-miR-466f-3p |
Kitl | 13,493 | 15 | up-regulated-miR-466f-3p | |
Fgf16 | 13,254 | 11 | up-regulated-miR-466f-3p | |
Grb2 | 11,784 | 15 | up-regulated-miR-466f-3p | |
Fgf1 | 11,640 | 11 | down-regulated-miR-342-5p |
Specific Keywords | |||
---|---|---|---|
Immune Responses | Immune Cells | Inflammation Signaling | |
Immune | T cell | Inflammation | JAK/Stat |
Cytotoxicity | B cell | Cytokine | CD3 |
Phagocytosis | Dendritic | Interleukin | CD4 |
Helper | NF-kappaB | CD8 | |
Macrophage | TGF-beta | ||
Myeloid |
Gene with 4 Degrees | Regulation by miRNA | Associated Keywords | |
---|---|---|---|
Nanog+F10-EVs | Rnf11 | up-regulated-miR-19a-3p | Immune, Cytokine, NF-kappaB, Tgf-beta |
iPS-EVs | Malt1 | up-regulated-miR-466f-3p | Immune, T cell, B cell, Helper cell |
Il1f6 | down-regulated-miR-30a-3p | T cell, Dendritic cell, Interleukin, NF-kappaB | |
Cmklr1 | down-regulated-miR-342-5p | Immune, Macrophage, Inflammation, NF-kappaB | |
Siglec1 | down-regulated-miR-342-5p | T cell, Macrophage, B cell, CD8 | |
Cd28 | down-regulated-miR-342-5p | Cytotoxicity, Immune, T cell, CD4 |
EVs | miRNA | Target Genes | Prediction By | Hub Gene Candidates |
---|---|---|---|---|
Nanog+F10-EVs/F10- EVs | up-regulated miRNAs (miR-18a-5p, miR-19a-3p, miR-3473b) | down-regulation of target genes | GD-based analysis | Trp53 |
Hif1a | ||||
Esr1 | ||||
Atm | ||||
SK-based analysis | Rnf11 | |||
down-regulated miRNAs (miR-706) | up-regulation of target genes | GD-based analysis | Cdkn1b | |
SK-based analysis | - | |||
iPS-EVs/Nanog+F10-EVs | up-regulated miRNAs (miR-466f-3p) | down-regulation of target genes | GD-based analysis | Ins1 |
Kitl | ||||
Fgf16 | ||||
Grb2 | ||||
SK-based analysis | Malt1 | |||
down-regulated miRNAs (miR-342-5p) | up-regulation of target genes | GD-based analysis | Fgf1 | |
SK-based analysis | Il1f6 | |||
Cmklr1 | ||||
Siglec1 | ||||
Cd28 |
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Khoo, C.S.M.; Henmi, T.; Saito, M. Comparative Study of Metastasis Suppression Effects of Extracellular Vesicles Derived from Anaplastic Cell Lines, Nanog-Overexpressing Melanoma, and Induced Pluripotent Stem Cells. Int. J. Mol. Sci. 2023, 24, 17206. https://doi.org/10.3390/ijms242417206
Khoo CSM, Henmi T, Saito M. Comparative Study of Metastasis Suppression Effects of Extracellular Vesicles Derived from Anaplastic Cell Lines, Nanog-Overexpressing Melanoma, and Induced Pluripotent Stem Cells. International Journal of Molecular Sciences. 2023; 24(24):17206. https://doi.org/10.3390/ijms242417206
Chicago/Turabian StyleKhoo, Celine Swee May, Takuya Henmi, and Mikako Saito. 2023. "Comparative Study of Metastasis Suppression Effects of Extracellular Vesicles Derived from Anaplastic Cell Lines, Nanog-Overexpressing Melanoma, and Induced Pluripotent Stem Cells" International Journal of Molecular Sciences 24, no. 24: 17206. https://doi.org/10.3390/ijms242417206