NLRP7 Promotes Choriocarcinoma Growth and Progression through the Establishment of an Immunosuppressive Microenvironment
Abstract
Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Human Study
2.1.1. Normal and Pathological Human Tissue and Sera
2.1.2. Immunohistochemistry of the Human Placentas
2.1.3. ELISA
2.2. Cell Culture
2.2.1. HTR8/SV Neo Cell Line Culture
2.2.2. JEG3 Cell Line Culture
2.2.3. JEG3 Luc Cell Line Preparation
2.2.4. JEG3-Sh-NLRP7 Cell Line Preparation
2.3. 2D Culture System
Proliferation Assay
2.4. 3D Culture System
2.4.1. Anchorage-Independent Spheroids Formation
2.4.2. Anchorage-Independent Colony Formation Assay in Soft Agar
2.5. RNA Isolation and Real-Time PCR Analysis
2.6. Western Blot Analysis
2.7. Gene Sequencing
2.8. In Vivo Studies
2.8.1. Experimental Groups
2.8.2. Histology and Immunohistochemistry of Mouse Tissues
2.8.3. Bioluminescence Imaging
2.9. RNA-Seq Analysis
2.10. Antibody Cytokine Arrays
2.11. Statistical Analysis
3. Results
3.1. NLRP7 Inflammasome Is Highly Expressed in CHM and CC
3.2. NLRP7 Inflammasome Is Not Active in the Human Choriocarcinoma Cell Line, JEG3
3.3. NLRP7 Knockdown in JEG3 Cells
3.4. NLRP7 Knockdown Decreases the Proliferation of JEG3 Tumor Formation In Vitro
3.5. Role of NLRP7 in CC Development and Progression in Two Preclinical Mouse Models
3.6. NLRP7 Knockdown Decreases CC Tumor Growth in the Mouse Placenta
3.7. NLRP7 Knockdown Decreases CC Tumor Growth in the Uterine Horn
3.8. NLRP7 Knockdown Rescued Maternal Adaptive Immune Response
3.9. A Proposed Model for NLRP7 Contribution to the Development of CC In Vivo
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Reynaud, D.; Abi Nahed, R.; Lemaitre, N.; Bolze, P.-A.; Traboulsi, W.; Sergent, F.; Battail, C.; Filhol, O.; Sapin, V.; Boufettal, H.; et al. NLRP7 Promotes Choriocarcinoma Growth and Progression through the Establishment of an Immunosuppressive Microenvironment. Cancers 2021, 13, 2999. https://doi.org/10.3390/cancers13122999
Reynaud D, Abi Nahed R, Lemaitre N, Bolze P-A, Traboulsi W, Sergent F, Battail C, Filhol O, Sapin V, Boufettal H, et al. NLRP7 Promotes Choriocarcinoma Growth and Progression through the Establishment of an Immunosuppressive Microenvironment. Cancers. 2021; 13(12):2999. https://doi.org/10.3390/cancers13122999
Chicago/Turabian StyleReynaud, Deborah, Roland Abi Nahed, Nicolas Lemaitre, Pierre-Adrien Bolze, Wael Traboulsi, Frederic Sergent, Christophe Battail, Odile Filhol, Vincent Sapin, Houssine Boufettal, and et al. 2021. "NLRP7 Promotes Choriocarcinoma Growth and Progression through the Establishment of an Immunosuppressive Microenvironment" Cancers 13, no. 12: 2999. https://doi.org/10.3390/cancers13122999
APA StyleReynaud, D., Abi Nahed, R., Lemaitre, N., Bolze, P.-A., Traboulsi, W., Sergent, F., Battail, C., Filhol, O., Sapin, V., Boufettal, H., Hoffmann, P., Aboussaouira, T., Murthi, P., Slim, R., Benharouga, M., & Alfaidy, N. (2021). NLRP7 Promotes Choriocarcinoma Growth and Progression through the Establishment of an Immunosuppressive Microenvironment. Cancers, 13(12), 2999. https://doi.org/10.3390/cancers13122999