Oncogenic Role of Secreted Engrailed Homeobox 2 (EN2) in Prostate Cancer
Abstract
:1. Introduction
2. Material and Methods
2.1. Patients and Samples
2.2. Datasets Analysis
2.3. EN2 Protein, Reagents, and Cell Lines
2.4. RNA Isolation, Reverse-Transcription, and Real-Time Quantitative PCR (qPCR)
2.5. Measurements of Cell Proliferation
2.6. Measurements of Migration Capacity
2.7. Measurement of Free Cytosolic Calcium Concentration ([Ca2+]i)
2.8. Microarray of Gene Expression Profile
2.9. Western Blot
2.10. Determination of PSA and EN2 Levels by ELISA
2.11. Statistical Analysis
3. Results
3.1. EN2 Is Overexpressed in Tissue and Urine Samples from PCa Patients Compared with Controls
3.2. EN2 Is Overexpressed, Secreted, and Modulates Aggressiveness Features in PCa Cells
3.3. EN2 Modulates Key Signaling Pathways and Molecular Targets in PCa Cells
4. Discussion
Supplementary Materials
Author Contributions
Financial Support
Acknowledgments
Conflicts of Interest
References
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Variable | Overall |
---|---|
Number of patients | 33 |
Age at diagnosis | |
Median (IQR) | 62 (58–66) |
BMI | |
Median (IQR) | 27.7 (25.8–31.3) |
PSA level, ng/mL | |
Median (IQR) | 6 (4.4–9.5) |
Gleason score in prostatectomy specimen (%) | |
6 | 9 (27.3) |
7 | 23 (69.7) |
8 | 1 (3) |
EE, n° (%) | 21 (63.6) |
PI, n° (%) | 28 (84.8) |
VI, n° (%) | 8 (24.2) |
Relative EN2 mRNA expression in FFPE piece a | |
Tumor tissue | |
Median (IQR) | 0.173 (0.002–1.473) |
Non-tumor adjacent tissue | |
Median (IQR) | 0.008 (0.000–0.477) |
Ratio tumor/non-tumor tissue | |
Median (IQR) | 3.451 (1.260–12.212) |
Relative EN1 mRNA expression in FFPE piece * | |
Tumor tissue | |
Median (IQR) | 0.723 (0.209–2.828) |
Non-tumor adjacent tissue | |
Median (IQR) | 0.421 (0.149–1.145) |
Ratio tumor/non-tumor tissue | |
Median (IQR) | 1.197 (0.325–3.249) |
Variable | Overall | Control | PCa |
---|---|---|---|
Patients | 30 | 7 | 23 |
Age at diagnosis | |||
Median (IQR) | 73 (64–79) | 67 (59–79) | 76 (67.0–80.0) |
PSA level, ng/mL | |||
Median (IQR) | - | - | 40 (22–70) |
Dyslipidemia (%) | 7 (23.3) | 2 (28.6) | 5 (21.7) |
Diabetes (%) | 8 (26.7) | 2 (28.6) | 6 (26.1) |
* BMI | |||
Median (IQR) | 27.19 (25.2–29.83) | 25.87 (24.50–34.24) | 27.44 (25.46–29.65) |
Gleason score | |||
=7 | - | - | 8 (34.8) |
>7 | - | - | 15 (65.2) |
EE (%) | - | - | 6 (26.1) |
PI (%) | - | - | 14 (60.9) |
# Metastasis (%) | - | - | 13 (56.5) |
N° samples (%) in whichEN2 was detected | 25 (83.3) | 3 (42.9) | 22 (95.7) |
ª Median(IQR) EN2 mRNAexpression | 445 (8–2265) | 0 (0–9) | 874 (173–2650) |
Variable | Control (n = 10) | Negative Biopsy (n = 10) | PCa (n = 24) | p-Value |
---|---|---|---|---|
Age, Years | ||||
Median (IQR) | 56 (52–61) | 56 (53–59) | 68 (59–71) | <0.01 |
Waist circumference, cm | ||||
Median (IQR) | 107 (99.5–111.8) | 103 (98.3–109) | 104.5 (100–111.5) | 0.88 |
BMI | ||||
Median (IQR) | 32.4 (27.5–33.1) | 30.1 (27.33–31.97) | 29 (26.67–30.48) | 0.15 |
PSA, ng/mL | ||||
Median (IQR) | 0.87 (0.6–1.6) | 3.6 (3.0–4.0) | 5.7 (4.6–9.7) | <0.01 |
DRE, abnormal (%) | 0 (0) | 0 (0) | 9 (37.5) | <0.01 |
Prostate Vol, cc | ||||
Median (IQR) | - | 37 (23.1–45.8) | 37 (25.59–52.5) | 0.55 |
1° Biopsy (%) | - | 9 (90) | 17 (70.8) | |
Gleason grade (%) | ||||
=6 | 6 (25) | |||
≥7 | 18 (75) | |||
Nº Pathologic cores | ||||
Median (IQR) | 2 (1–4) | |||
EN2 Urine | ||||
Median (IQR) | 0 (0–0.21) | 0.02 (0.00–0.30) | 0.19 (0.01–0.43) | 0.05 |
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Gómez-Gómez, E.; Jiménez-Vacas, J.M.; Pedraza-Arévalo, S.; López-López, F.; Herrero-Aguayo, V.; Hormaechea-Agulla, D.; Valero-Rosa, J.; Ibáñez-Costa, A.; León-González, A.J.; Sánchez-Sánchez, R.; et al. Oncogenic Role of Secreted Engrailed Homeobox 2 (EN2) in Prostate Cancer. J. Clin. Med. 2019, 8, 1400. https://doi.org/10.3390/jcm8091400
Gómez-Gómez E, Jiménez-Vacas JM, Pedraza-Arévalo S, López-López F, Herrero-Aguayo V, Hormaechea-Agulla D, Valero-Rosa J, Ibáñez-Costa A, León-González AJ, Sánchez-Sánchez R, et al. Oncogenic Role of Secreted Engrailed Homeobox 2 (EN2) in Prostate Cancer. Journal of Clinical Medicine. 2019; 8(9):1400. https://doi.org/10.3390/jcm8091400
Chicago/Turabian StyleGómez-Gómez, Enrique, Juan M. Jiménez-Vacas, Sergio Pedraza-Arévalo, Fernando López-López, Vicente Herrero-Aguayo, Daniel Hormaechea-Agulla, José Valero-Rosa, Alejandro Ibáñez-Costa, Antonio J. León-González, Rafael Sánchez-Sánchez, and et al. 2019. "Oncogenic Role of Secreted Engrailed Homeobox 2 (EN2) in Prostate Cancer" Journal of Clinical Medicine 8, no. 9: 1400. https://doi.org/10.3390/jcm8091400