Inhibiting P2Y12 in Macrophages Induces Endoplasmic Reticulum Stress and Promotes an Anti-Tumoral Phenotype
Abstract
:1. Introduction
2. Results
2.1. The P2Y12 Receptor Is Expressed in Mouse Models for Liver Cirrhosis and Hepatocellular Carcinoma
2.2. Expression of P2Y12 Is Located in the Stroma of Patients with Hepatocellular Carcinoma
2.3. Inhibition of the P2Y12 Receptor Decreases Viability of Macrophages
2.4. Ticagrelor Enhances Phagocytosis of Tumor Cells
2.5. Expression of ER Stress Genes Is Increased after P2Y12 Inhibition with Ticagrelor
3. Discussion
4. Materials and Methods
4.1. Animal Models
4.2. Cell Culture and Reagents
4.3. Differentiation of THP1 Cells
4.4. Proliferation
4.5. Phagocytosis Assay
4.6. Immunohistochemistry
4.7. SDS-PAGE and Western Blot
4.8. Flow Cytometry
4.9. RT-qPCR
4.10. Human Protein Atlas
4.11. Live/Dead Cell Double Staining Kit
4.12. Statistics
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
ADP | Adenosine diphosphate |
ATF6 | Activating transcription factor 6 |
ATP | Adenosine triphosphate |
BIP | Binding immunoglobulin protein |
CCL4 | Carbon tetrachloride |
DEN | Diethylnitrosamine |
DMEM | Dulbecco modified eagle medium |
FPKM | Fragments Per Kilobase of exon per Million |
HCC | Hepatocellular carcinoma |
IRE1α | Inositol-requiring enzyme alpha |
IL | Interleukin |
MAPK | Mitogen-activated protein kinase |
PBS | Phosphate buffered saline |
PERK | Protein kinase R-like endoplasmic reticulum kinase |
Tudca | Tauroursodeoxycholic acid |
UPR | Unfolded protein response |
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Target (Species) | Forward | Reverse |
---|---|---|
P2ry12 (mouse) | ACGAGTTTGTGAAGGCACGA | AGACTCACGACTTTCTTCTTTGGA |
P2ry12 (human) | TTTGCCCGAATTCCTTACAC | ATTGGGGCACTTCAGCATAC |
18S (human) | AGTCCCTGCCCTTTGTACACA | GATCCGAGGGCCTCACTAAAC |
CHOP (human) | CATCACCACACCTGAAAGCA | TCAGCTGCCATCTCTGCA |
XBP1-spliced (human) | AGACAGCGCTTGGGGATGGAT | CCTGCACCTGCTGCGGACTC |
BIP (human) | GAAAGAAGGTTACCCATGCAGT | CAGGCCATAAGCAATAGCAGC |
GAPDH (human) | GGAGCGAGATCCCTCCAAAAT | GGCTGTTGTCATACTTCTCATGG |
GAPDH (mouse) | AATGAAGGGGTCGTTGATG | GGTGAAGGTCGGTGTGAAC |
TBP1 (human) | AGTGACCCAGCATCACTGTTT | GGCAAACCAGAAACCCTTGC |
ATF4 (human) | TCAGACAGTACCAACGCTTATGC | GTTGTACCACAGTAGGCTGAGA |
ATF6 (human) | GTTGTACCACAGTAGGCTGAGA | TGGCCAAGCACTTCAAACCT |
IRE1 (human) | TAGTCAGTTCTGCGTCCGCT | TTCCAAAAATCCCGAGGCCG |
PERK (human) | ACGATGAGACAGAGTTGCGA | GCGCGTAAACAAGTTGCCT |
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Pavlović, N.; Kopsida, M.; Gerwins, P.; Heindryckx, F. Inhibiting P2Y12 in Macrophages Induces Endoplasmic Reticulum Stress and Promotes an Anti-Tumoral Phenotype. Int. J. Mol. Sci. 2020, 21, 8177. https://doi.org/10.3390/ijms21218177
Pavlović N, Kopsida M, Gerwins P, Heindryckx F. Inhibiting P2Y12 in Macrophages Induces Endoplasmic Reticulum Stress and Promotes an Anti-Tumoral Phenotype. International Journal of Molecular Sciences. 2020; 21(21):8177. https://doi.org/10.3390/ijms21218177
Chicago/Turabian StylePavlović, Nataša, Maria Kopsida, Pär Gerwins, and Femke Heindryckx. 2020. "Inhibiting P2Y12 in Macrophages Induces Endoplasmic Reticulum Stress and Promotes an Anti-Tumoral Phenotype" International Journal of Molecular Sciences 21, no. 21: 8177. https://doi.org/10.3390/ijms21218177