Insights into Intra-Tumoral Heterogeneity: Transcriptional Profiling of Chemoresistant MPM Cell Subpopulations Reveals Involvement of NFkB and DNA Repair Pathways and Contributes a Prognostic Signature
Abstract
:1. Introduction
2. Results
2.1. ALDH1A3 Expression May Stratify MPM Patients
2.2. Gene Expression Profile of Purified ALDHbright and ALDHlow Cells, at Steady State and after Butein Treatment
2.3. Analyzing the Gene Expression Profile of ALDHbright and ALDHlow Cell Subpopulations
2.4. The MPM ALDHbright Cells Exhibited Enrichment for NFkB-Related Pathways
2.5. The mRNAs Downregulated More Deeply in the ALDHbright Cells by Butein Were Related to DNA Replication and Repair Functions
2.6. mRNAs Enriched in the ALDHbright Cells May Have Prognostic Potential
3. Discussion
4. Materials and Methods
4.1. Cell Lines and Treatments
4.2. ALDH Detection
4.3. FACS-Based Purification of ALDHbright and ALDHlow Cells
4.4. RNA Extraction and Analysis
4.5. Q-RTPCR
4.6. Microarray Analysis
4.7. Differential Gene Expression Statistical Analysis
4.8. Validation of the mRNA Expression
4.9. Principal Component Analysis (PCA)
4.10. Pathway Enrichment Analysis
4.11. Identification of a Prognostic MPM Signature
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
ALDH | Aldehyde Dehydrogenase |
MPM | Malignant Pleural Mesothelioma |
ELISA | Enzyme-linked immunosorbent assay |
NFkB | nuclear factor of kappa light polypeptide gene enhancer in B-cells |
CSC | Cancer Stem Cell |
EMT | Epithelial to Mesenchymal Transition |
FACS | Fluorescence Activated Cell Sorting |
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Cioce, M.; Sacconi, A.; Pass, H.I.; Canino, C.; Strano, S.; Blandino, G.; Fazio, V.M. Insights into Intra-Tumoral Heterogeneity: Transcriptional Profiling of Chemoresistant MPM Cell Subpopulations Reveals Involvement of NFkB and DNA Repair Pathways and Contributes a Prognostic Signature. Int. J. Mol. Sci. 2021, 22, 12071. https://doi.org/10.3390/ijms222112071
Cioce M, Sacconi A, Pass HI, Canino C, Strano S, Blandino G, Fazio VM. Insights into Intra-Tumoral Heterogeneity: Transcriptional Profiling of Chemoresistant MPM Cell Subpopulations Reveals Involvement of NFkB and DNA Repair Pathways and Contributes a Prognostic Signature. International Journal of Molecular Sciences. 2021; 22(21):12071. https://doi.org/10.3390/ijms222112071
Chicago/Turabian StyleCioce, Mario, Andrea Sacconi, Harvey I. Pass, Claudia Canino, Sabrina Strano, Giovanni Blandino, and Vito Michele Fazio. 2021. "Insights into Intra-Tumoral Heterogeneity: Transcriptional Profiling of Chemoresistant MPM Cell Subpopulations Reveals Involvement of NFkB and DNA Repair Pathways and Contributes a Prognostic Signature" International Journal of Molecular Sciences 22, no. 21: 12071. https://doi.org/10.3390/ijms222112071