Extracellular Vesicles-Mediated Transfer of miRNA Let-7b from PC3 Cells to Macrophages
Abstract
:1. Introduction
2. Materials and Methods
2.1. Material
2.2. Cell Culture and Treatments
2.3. Isolation of Extracellular Vesicles (EVs) and Preparation of Conditioned Medium Depleted of EVs (CM-EVs)
2.4. Extracellular Vesicles Staining and Fluorescence Microscopy
2.5. RNA Isolation, cDNA Synthesis, PCR and QPCR
2.6. Statistical Analysis
3. Results
3.1. Expression of Endogenous miRNA Let-7b in Cancerous and Noncancerous Prostate Cell Lines
3.2. Differential Let-7b miRNA Content in EVs Compared to Prostate Cells
3.3. Evaluation of miRNA Let-7b in THP-1 Cells Treated with PC3-Derived EVs
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Gene | Forward (F) | Reverse (R) |
---|---|---|
Pre-miRNA Let-7b | 5′ TGAGGTAGTAGGTTGTGTGG 3′ | 5′ CAGGGAAGGCAGTAGGTTG 3′ |
nested pre-miRNA Let-7b | 5′ TTTCAGGGCAGTGATGTTGC 3′ | |
miRNA Let-7b | 5′ TGAGGTAGTAGGTTGTGTGGTT 3′ | 5′ GTGCAGGGTCCGAGGT 3′ |
IL-12 p40 | 5′ CGGTCATCTGCCGCAAA 3′: | 5′ TGCCCATTCGCTCCAAGA 3′ |
IL-10 | 5′ CGAGATGCCTTCAGCAGAGT 3′ | 5′ CGCCTTGATGTCTGGGTCTT 3′ |
stem loop miRNA Let-7b | 5′ GTCGTATCCAGTGCAGGGTCCGAGGTGCACTGGATACGACCACCCACCAACCAC 3′ |
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Costanzi, E.; Romani, R.; Scarpelli, P.; Bellezza, I. Extracellular Vesicles-Mediated Transfer of miRNA Let-7b from PC3 Cells to Macrophages. Genes 2020, 11, 1495. https://doi.org/10.3390/genes11121495
Costanzi E, Romani R, Scarpelli P, Bellezza I. Extracellular Vesicles-Mediated Transfer of miRNA Let-7b from PC3 Cells to Macrophages. Genes. 2020; 11(12):1495. https://doi.org/10.3390/genes11121495
Chicago/Turabian StyleCostanzi, Egidia, Rita Romani, Paolo Scarpelli, and Ilaria Bellezza. 2020. "Extracellular Vesicles-Mediated Transfer of miRNA Let-7b from PC3 Cells to Macrophages" Genes 11, no. 12: 1495. https://doi.org/10.3390/genes11121495