Silencing of circCRIM1 Drives IGF2BP1-Mediated NSCLC Immune Evasion
Abstract
:1. Introduction
2. Materials and Methods
2.1. Tissue Specimen Collection
2.2. Cell Culture and Treatment
2.3. RNA Extraction and qRT-PCR
2.4. RNase R Treatment
2.5. Actinomycin D Treatment and RNA Stability Assay
2.6. Nuclear and Cytoplasmic Extraction
2.7. Vector Construction and Cell Transfection
2.8. Isolation and Activation of CD8 + T Cells and NK Cells
2.9. In Vitro Co-Culture Assay
2.10. RNA Pull-Down Assays and Mass Spectrometry Analysis
2.11. Western Blot and RNA Immunoprecipitation (RIP)
2.12. Fluorescent In Situ Hybridization (FISH) and Immunofluorescence
2.13. Dual-Luciferase Reporter Assay
2.14. In Vivo Tumorigenesis Assays
2.15. Flow Cytometry Analysis
2.16. Immunohistochemistry and Terminal Deoxyribonucleotide Transferase-Mediated Nick-End Labeling (TUNEL) Analyses
2.17. RNA Sequencing
2.18. Cell Counting Kit-8 (CCK-8), Colony Formation and CFSE Proliferation Assays
2.19. Transwell Migration and Apoptosis Assays
2.20. Statistical Analysis
3. Results
3.1. Identification of circCRIM1 as a Candidate NSCLC Suppressor
3.2. Characterization of circCRIM1 in NSCLC Cells
3.3. CircCRIM1 Modulation Alters NSCLC Cells Immune Evasion In Vitro
3.4. CircCRIM1–IGF2BP1 Interaction in the Cytoplasm of NSCLC
3.5. CircCRIM1 Destabilized HLA-F mRNA via Competing Interaction with IGF2BP1 in NSCLC
3.6. CircCRIM1 Is a Potential Therapeutic Target to Inhibit NSCLC Progression
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Peng, W.; Ye, L.; Xue, Q.; Wei, X.; Wang, Z.; Xiang, X.; Zhang, S.; Zhang, P.; Wang, H.; Zhou, Q. Silencing of circCRIM1 Drives IGF2BP1-Mediated NSCLC Immune Evasion. Cells 2023, 12, 273. https://doi.org/10.3390/cells12020273
Peng W, Ye L, Xue Q, Wei X, Wang Z, Xiang X, Zhang S, Zhang P, Wang H, Zhou Q. Silencing of circCRIM1 Drives IGF2BP1-Mediated NSCLC Immune Evasion. Cells. 2023; 12(2):273. https://doi.org/10.3390/cells12020273
Chicago/Turabian StylePeng, Wenbei, Linlin Ye, Qianqian Xue, Xiaoshan Wei, Zihao Wang, Xuan Xiang, Siyu Zhang, Pei Zhang, Haolei Wang, and Qiong Zhou. 2023. "Silencing of circCRIM1 Drives IGF2BP1-Mediated NSCLC Immune Evasion" Cells 12, no. 2: 273. https://doi.org/10.3390/cells12020273
APA StylePeng, W., Ye, L., Xue, Q., Wei, X., Wang, Z., Xiang, X., Zhang, S., Zhang, P., Wang, H., & Zhou, Q. (2023). Silencing of circCRIM1 Drives IGF2BP1-Mediated NSCLC Immune Evasion. Cells, 12(2), 273. https://doi.org/10.3390/cells12020273