Loss of Two-Pore Channel 2 (TPC2) Expression Increases the Metastatic Traits of Melanoma Cells by a Mechanism Involving the Hippo Signalling Pathway and Store-Operated Calcium Entry
Abstract
1. Introduction
2. Results
2.1. Association between TPC2 Expression and the Prognosis of Melanoma Patients
2.2. TPC2 Depletion Impairs Human Melanoma Cell Adhesion Mediated by α2β1 Integrin Receptor
2.3. TPC2 Inhibition in Human Melanoma Cells Increases Their Invasion Capability
2.4. TPC2 and HIPPO Pathway Connection in CHL1 and MeWo Metastatic Cell Lines
2.5. PD-L1 Induced by INTERFERON-γ Is Significantly Higher in CHL1 TPC2 KO Cells
2.6. TPC2 Regulates YAP/TAZ Activity Modulating ORAI 1 Channel Expression
2.7. TPC2 Is Overexpressed while YAP/TAZ Target Genes Are Down Regulated after Vemurafenib Treatment in A375 Melanoma Cells
3. Discussion
4. Materials and Methods
4.1. Cell Lines
4.2. Western Blot Analysis
4.3. CRISPR Design
4.4. Cell Transfection
4.5. Adhesion Assay
4.6. Invasion Assay
4.7. Flow Cytometry and Cell Cycle Analysis
4.8. Bioinformatic Analysis
4.9. Quantitative Real-Time PCR
4.10. Gelatin Zymography for Matrix Metalloproteinases (MMPs) Detection
4.11. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Gene | Forward | Reverse |
---|---|---|
CTGF | AGGAGTGGGTGTGTGACGA | CCAGGCAGTTGGCTCTAATC |
CYR61 | CCTTGTGGACAGCCAGTGTA | ACTTGGGCCGGTATTTCTTC |
ANKRD1 | AGTAGAGGAACTGGTCACTGG | TGGGCTAGAAGTGTCTTCAGAT |
YAP | GCACCTCTGTGTTTTAAGGGTCT | CAACTTTTGCCCTCCTCCAA |
TAZ(WWTR1) | GGCTGGGAGATGACCTTCAC | CTGAGTGGGGTGGTTCTGCT |
GAPDH | GGAGCGAGATCCCTCCAAAAT | GGCTGTTGTCATACTTCTCATGG |
STIM | ATCTCACAGCTCATGGTATGCTCC | GGAAGGTGCCAAAGAGTGTGTTTC |
ORAI | TACTTGAGCCGCGCCAAGCTTAAA | ACCGAGTTGAGATTGTGCACGTTG |
PKCβII | GACCAAACACCCAGGCAAAC | GATGGCGGGTGAAAAATCGG |
INF2 | CACATCCAACGTGATGGTGAAG | GGAGAGCTCGTTCATGACAATG |
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D’Amore, A.; Hanbashi, A.A.; Di Agostino, S.; Palombi, F.; Sacconi, A.; Voruganti, A.; Taggi, M.; Canipari, R.; Blandino, G.; Parrington, J.; et al. Loss of Two-Pore Channel 2 (TPC2) Expression Increases the Metastatic Traits of Melanoma Cells by a Mechanism Involving the Hippo Signalling Pathway and Store-Operated Calcium Entry. Cancers 2020, 12, 2391. https://doi.org/10.3390/cancers12092391
D’Amore A, Hanbashi AA, Di Agostino S, Palombi F, Sacconi A, Voruganti A, Taggi M, Canipari R, Blandino G, Parrington J, et al. Loss of Two-Pore Channel 2 (TPC2) Expression Increases the Metastatic Traits of Melanoma Cells by a Mechanism Involving the Hippo Signalling Pathway and Store-Operated Calcium Entry. Cancers. 2020; 12(9):2391. https://doi.org/10.3390/cancers12092391
Chicago/Turabian StyleD’Amore, Antonella, Ali Ahmed Hanbashi, Silvia Di Agostino, Fioretta Palombi, Andrea Sacconi, Aniruddha Voruganti, Marilena Taggi, Rita Canipari, Giovanni Blandino, John Parrington, and et al. 2020. "Loss of Two-Pore Channel 2 (TPC2) Expression Increases the Metastatic Traits of Melanoma Cells by a Mechanism Involving the Hippo Signalling Pathway and Store-Operated Calcium Entry" Cancers 12, no. 9: 2391. https://doi.org/10.3390/cancers12092391
APA StyleD’Amore, A., Hanbashi, A. A., Di Agostino, S., Palombi, F., Sacconi, A., Voruganti, A., Taggi, M., Canipari, R., Blandino, G., Parrington, J., & Filippini, A. (2020). Loss of Two-Pore Channel 2 (TPC2) Expression Increases the Metastatic Traits of Melanoma Cells by a Mechanism Involving the Hippo Signalling Pathway and Store-Operated Calcium Entry. Cancers, 12(9), 2391. https://doi.org/10.3390/cancers12092391