Six-Gene Signature for Differential Diagnosis and Therapeutic Decisions in Non-Small-Cell Lung Cancer—A Validation Study
Abstract
:1. Introduction
2. Results
2.1. Patients’ Characteristics
2.2. Differences in the Expression Profiles of Validated Genes between the Histological Subtypes of NSCLC (SCC vs. ADC)
2.3. Relationships between Expression Levels of Validated Genes and Clinicopathological Characteristics of NSCLC Patients
2.4. Visualizing Decision Boundary for Logistic Regression Models of Individual Validated Genes
2.5. Gene Expression Value in NSCLC Subtyping Based on Logistic Regression and Gradient Boosting Decision Tree Models
2.6. Prognostic Significance of Validated Gene Expression in NSCLC Patients
2.7. Gene Ontology Analyses
3. Discussion
4. Materials and Methods
4.1. Patients and Samples
4.2. Histopathological Diagnosis
4.3. RNA Extraction and Quality Control
4.4. Quantitative Real-Time PCR Analysis
4.5. Statistical Analysis
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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Gender | N | % |
---|---|---|
Male | 107 | 76.43 |
Female | 33 | 23.57 |
Age at Diagnosis (years) | ||
Range | 40–81 | |
Mean | 63.33 | |
Histological Type | ||
Squamous cell carcinoma | 79 | 56.42 |
Adenocarcinoma | 61 | 43.57 |
TNM Stage | ||
IA | 33 | 23.57 |
IB | 32 | 22.86 |
IIA | 28 | 20.0 |
IIB | 38 | 27.14 |
IIIA | 8 | 5.71 |
Information not available | 1 | 0.7 |
Recurrence | ||
Present | 54 | 38.57 |
Absent | 82 | 58.57 |
Information not available | 4 | 2.86 |
Genes ∆Ct SCC | Genes ∆Ct ADC | Expression Profile | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
N | Mean | SD | 25% | 75% | N | Mean | SD | 25% | 75% | p-Value | SCC vs. ADC | |
MIR-205HG | 81 | 14.808 | 2.769 | 13.226 | 15.362 | 59 | 20.099 | 3.426 | 17.940 | 22.425 | <0.001 | up |
SERPINB5 | 81 | 14.850 | 2.990 | 12.725 | 15.977 | 59 | 20.305 | 3.640 | 18.546 | 22.048 | <0.001 | up |
KRT6C | 81 | 16.697 | 3.104 | 14.670 | 18.749 | 59 | 18.422 | 3.041 | 16.061 | 20.628 | 0.003381 | up |
KRT6A | 81 | 11.574 | 3.145 | 9.580 | 13.001 | 59 | 16.578 | 2.858 | 14.772 | 19.071 | <0.001 | up |
KRT5 | 81 | 11.968 | 3.492 | 9.543 | 12.405 | 59 | 19.383 | 3.369 | 17.277 | 21.823 | <0.001 | up |
DSG | 81 | 14.864 | 4.012 | 12.172 | 16.386 | 59 | 23.050 | 3.559 | 21.261 | 25.255 | <0.001 | up |
GO Category | GO Term Id | GO Term Label | p-Value | Genes Involved |
---|---|---|---|---|
Cellular component | GO:0001533 | cornified envelope | 8.86 × 10−5 | DSG3, SERPINB5 |
GO:0045095 | keratin filament | 1.10 × 10−6 | KRT6A, KRT5, KRT6C | |
GO:0005882 | intermediate filament | 1.35 × 10−5 | KRT6A | |
GO:0045111 | intermediate filament cytoskeleton | 2.09 × 10−5 | KRT6A, KRT5, KRT6C | |
GO:0005615 | extracellular space | 0.00011 | DSG3, KRT6A, KRT5, KRT6C, SERPINB5 | |
Biological process | GO:0045109 | intermediate filament organization | 4.99 × 10−7 | KRT6A, KRT5, KRT6C |
GO:0045104 | intermediate filament cytoskeleton organization | 1.04 × 10−6 | KRT6A, KRT5, KRT6C | |
GO:0045103 | intermediate filament-based process | 1.07 × 10−6 | KRT6A, KRT5, KRT6C | |
GO:0031424 | keratinization | 6.99 × 10−7 | KRT6A, KRT5, KRT6C | |
GO:0030216 | keratinocyte differentiation | 3.67 × 10−6 | KRT6A, KRT5, KRT6C | |
GO:0009913 | epidermal cell differentiation | 1.10 × 10−5 | KRT6A, KRT5, KRT6C | |
Molecular function | GO:0030280 | structural constituent of skin epidermis | 6.77 × 10−8 | KRT6A, KRT5, KRT6C |
RefSeq | Gene Name, Abbreviation | Assay IDs from Applied Biosystems (Primers + Probe) |
---|---|---|
NM_000424.3 | Keratin 5 (KRT5) | Hs00361185_m1 |
NM_005554.3 | Keratin 6A (KRT6A) | Hs01699178_g1 |
NM_002639.4 XM_006722483.2 | Serpin Family B Member 5 (SERPINB5) | Hs00985285_m1 |
NM_001104548.1 XM_017002058.1 | MIR205 Host Gene (MIR205HG) | Hs03405498_m1 |
NM_173086.4 | Keratin 6C (KRT6C) | Hs00752476_s1 |
NM_001944.2 XM_011525850.1 | Desmoglein 3 (DSG3) | Hs00951897_m1 |
X03205.1 | 18S rRNA (Reference Gene) | Hs99999901_s1 |
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Charkiewicz, R.; Sulewska, A.; Karabowicz, P.; Lapuc, G.; Charkiewicz, A.; Kraska, M.; Pancewicz, J.; Lukasik, M.; Kozlowski, M.; Stec, R.; et al. Six-Gene Signature for Differential Diagnosis and Therapeutic Decisions in Non-Small-Cell Lung Cancer—A Validation Study. Int. J. Mol. Sci. 2024, 25, 3607. https://doi.org/10.3390/ijms25073607
Charkiewicz R, Sulewska A, Karabowicz P, Lapuc G, Charkiewicz A, Kraska M, Pancewicz J, Lukasik M, Kozlowski M, Stec R, et al. Six-Gene Signature for Differential Diagnosis and Therapeutic Decisions in Non-Small-Cell Lung Cancer—A Validation Study. International Journal of Molecular Sciences. 2024; 25(7):3607. https://doi.org/10.3390/ijms25073607
Chicago/Turabian StyleCharkiewicz, Radoslaw, Anetta Sulewska, Piotr Karabowicz, Grzegorz Lapuc, Alicja Charkiewicz, Marcin Kraska, Joanna Pancewicz, Malgorzata Lukasik, Miroslaw Kozlowski, Rafal Stec, and et al. 2024. "Six-Gene Signature for Differential Diagnosis and Therapeutic Decisions in Non-Small-Cell Lung Cancer—A Validation Study" International Journal of Molecular Sciences 25, no. 7: 3607. https://doi.org/10.3390/ijms25073607