Ras-Related Protein Rab-32 and Thrombospondin 1 Confer Resistance to the EGFR Tyrosine Kinase Inhibitor Osimertinib by Activating Focal Adhesion Kinase in Non-Small Cell Lung Cancer
Abstract
:Simple Summary
Abstract
1. Introduction
2. Material and Methods
2.1. Cell Culture
2.2. Chemicals and Antibodies
2.3. Cell Viability Assay
2.4. Wound Healing Assay
2.5. Western Blot
2.6. Transcriptome Analysis
2.7. Functional Enrichment Analysis and TCGA Co-Expression Analysis
2.8. siRNA Library Screen and Knockdown
2.9. RNA Isolation and Quantitative Real-Time PCR (RT-qPCR)
2.10. Patient Cohort and Sample Preparation
2.11. Exosome RNA Extraction
2.12. Ethics Statement
2.13. Statistical Analysis
3. Results
3.1. Characterization of Parental and Osimertinib-Resistant NSCLC Cell Lines
3.2. Osimertinib-Resistant Cells Acquired an Enhanced EMT Phenotype
3.3. Transcriptome Profiling of Osimertinib-Resistant NSCLC Cells
3.4. siRNA Library Screening Analyses of Upregulated Genes in Osimertinib-Resistant Cells
3.5. RAB32 and THBS1 Were Elevated in Expression in NSCLC Patients with Disease Progression on Osimertinib
3.6. RAB32 and THBS1 Are Mechanistically Linked to Focal Adhesion Kinase
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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HCC827OR/P | NCI-H1975OR/P | ||||||||
---|---|---|---|---|---|---|---|---|---|
Gene Symbol | Description | p-Value | FDR | Fold Change | Cell Viability% in HCC827OR/P | p-Value | FDR | Fold Change | Cell Viability % in NCI-H1975OR/P |
IGFBP7 | Insulin-like growth factor-binding protein 7 | 3.17 × 10−11 | 0 | 88.52 | 43.12% | 1.31 × 10−9 | 2.55 × 10−5 | 28.58 | 56.81% |
CADM1 | Cell adhesion molecule 1 | 0.0000704 | 0.0075 | 5.15 | 60.90% | 4.63 × 10−8 | 0.0002 | 24.01 | 66.24% |
COL5A1 | Collagen, type V, alpha 1 | 4.85 × 10−9 | 0 | 17.97 | 61.22% | 8.95 × 10−7 | 0.0017 | 6.5 | 34.08% |
AKT3 | v-akt Murine thymoma viral oncogene homolog 3 | 0.0000884 | 0.0087 | 5.59 | 76.78% | 7.43 × 10−5 | 0.0225 | 5.77 | 66.20% |
PTPRM | Protein tyrosine phosphatase, receptor type, M | 0 | 0.00006 | 10.6 | 61.58% | 5.03 × 10−6 | 0.0046 | 5.43 | 65.46% |
BICC1 | BicC family RNA-binding protein 1 | 0.0002 | 0.0129 | 4.89 | 51.75% | 9.91 × 10−5 | 0.0259 | 5.3 | 49.36% |
RAB32 | RAB32, member RAS oncogene family | 0.0001 | 0.0115 | 3.47 | 49.18% | 0.0002 | 0.035 | 3.32 | 58.71% |
THBS1 | Thrombospondin 1 | 0 | 0.00001 | 10.24 | 69.59% | 4.88 × 10−5 | 0.0177 | 3.18 | 70.81% |
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Kosibaty, Z.; Brustugun, O.T.; Zwicky Eide, I.J.; Tsakonas, G.; Grundberg, O.; De Petris, L.; McGowan, M.; Hydbring, P.; Ekman, S. Ras-Related Protein Rab-32 and Thrombospondin 1 Confer Resistance to the EGFR Tyrosine Kinase Inhibitor Osimertinib by Activating Focal Adhesion Kinase in Non-Small Cell Lung Cancer. Cancers 2022, 14, 3430. https://doi.org/10.3390/cancers14143430
Kosibaty Z, Brustugun OT, Zwicky Eide IJ, Tsakonas G, Grundberg O, De Petris L, McGowan M, Hydbring P, Ekman S. Ras-Related Protein Rab-32 and Thrombospondin 1 Confer Resistance to the EGFR Tyrosine Kinase Inhibitor Osimertinib by Activating Focal Adhesion Kinase in Non-Small Cell Lung Cancer. Cancers. 2022; 14(14):3430. https://doi.org/10.3390/cancers14143430
Chicago/Turabian StyleKosibaty, Zeinab, Odd Terje Brustugun, Inger Johanne Zwicky Eide, Georgios Tsakonas, Oscar Grundberg, Luigi De Petris, Marc McGowan, Per Hydbring, and Simon Ekman. 2022. "Ras-Related Protein Rab-32 and Thrombospondin 1 Confer Resistance to the EGFR Tyrosine Kinase Inhibitor Osimertinib by Activating Focal Adhesion Kinase in Non-Small Cell Lung Cancer" Cancers 14, no. 14: 3430. https://doi.org/10.3390/cancers14143430