Novel Gastric Cancer Stem Cell-Related Marker LINGO2 Is Associated with Cancer Cell Phenotype and Patient Outcome
Abstract
:1. Introduction
2. Results
2.1. LINGO2 is Differentially Expressed in Gastric Cancer Sphere Cells
2.2. Increase in LINGO2 Expression Elevates Cancer Stem Cell Characteristics
2.3. Silencing LINGO2 Reduces Cell Proliferation and Motility
2.4. LINGO2 Alters Secretion of Matrix Metallopeptidases and Angiogenic Factors
2.5. Clinical Classification of Gastric Cancer Patients Based on LINGO2 Expression
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Sphere-Formation Assay
4.3. Microarray Analysis
4.4. Knockdown of LINGO2 by shRNA
4.5. Semiquantitative Reverse-Transcription Polymerase Chain Reaction (RT-PCR)
4.6. Western Blotting
4.7. Immunofluorescence Staining
4.8. Cell Proliferation Assays
4.9. Migration and Invasion Assays
4.10. Wound-Healing Assays
4.11. Clonogeinc Assays
4.12. Gel Zymography
4.13. Flow Cytometry
4.14. Tumorigenesis Assays
4.15. Tube Formation Assay
4.16. Patients
4.17. Immunohistochemistry
4.18. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
LINGO2 | leucine-rich repeat and immunoglobulin-like domain-containing nogo receptor-interacting protein 2 |
CSC | cancer stem cells |
EMT | epithelial to mesenchymal transition |
TMA | tissue microarray analysis |
SPEM | spasmolytic polypeptide-expressing metaplasia |
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Total n = 103 | LINGO2 Weak n = 75 | LINGO2 Strong n = 28 | p | |
---|---|---|---|---|
Mean age, years (SD) | 57.9 (10.7) | 58.3 (10.5) | 56.9 (11.3) | 0.549 |
Sex (%) | ||||
Male | 67 (65.0) | 50 (66.7) | 17 (60.7) | 0.573 |
Female | 36 (35.0) | 25 (33.3) | 11 (39.3) | |
CEA, ng/mL (SD) | 8.0 (26.3) | 8.0 (26.3) | 8.1 (27.1) | 0.992 |
Median OS, months (range) | 96.9 (2.0–220.8) | 121.2 (2.0–220.8) | 25.5 (5.3–219.3) | 0.012 |
pT stage (%) | 0.011 | |||
T1 | 35 (34.0) | 29 (38.7) | 6 (21.4) | |
T2 | 23 (22.3) | 20 (26.7) | 3 (10.7) | |
T3 | 43 (41.7) | 24 (32.0) | 19 (67.9) | |
T4 | 2 (1.9) | 2 (2.7) | 0 (0.0) | |
T1/2 | 58 (56.3) | 49 (65.3) | 9 (32.1) | 0.003 |
T3/4 | 45 (43.7) | 26 (34.7) | 19 (67.9) | |
pN stage (%) | 0.005 | |||
N0 | 45 (43.7) | 39 (52.0) | 6 (21.4) | |
N1 | 27 (26.2) | 19 (25.3) | 8 (28.6) | |
N2 | 24 (23.3) | 15 (20.0) | 9 (32.1) | |
N3 | 7 (6.8) | 2 (2.7) | 5 (17.9) | |
N0/1 | 72 (69.9) | 58 (77.3) | 14 (50.0) | 0.007 |
N2/3 | 31 (30.1) | 17 (22.7) | 14 (50.0) | |
Stage (%) | 0.015 | |||
IA | 26 (25.2) | 23 (30.7) | 3 (10.7) | |
IB | 28 (27.2) | 22 (29.3) | 6 (21.4) | |
IIB | 22 (21.4) | 17 (22.7) | 5 (17.9) | |
IIIA | 18 (17.5) | 9 (12.0) | 9 (32.1) | |
IIIB | 9 (8.7) | 4 (5.3) | 5 (17.9) | |
I/II | 76 (73.8) | 62 (82.7) | 14 (50.0) | 0.001 |
III | 27 (26.2) | 13 (17.3) | 14 (50.0) |
Univariate | Multivariate | |||
---|---|---|---|---|
Hazard Ratio (95% CI) | p | Hazard Ratio (95% CI) | p | |
LINGO2strong (vs. LINGO2 weak) | 1.939 (1.146–3.279) | 0.014 | 1.245 (0.674–2.301) | 0.484 |
Older Age | 1.053 (1.025–1.082) | <0.001 | 1.053 (1.023–1.083) | <0.001 |
Male sex (vs. female) | 1.160 (0.689–1.956) | 0.576 | 1.433 (0.805–2.551) | 0.222 |
Higher initial CEA | 1.007 (0.998–1.016) | 0.141 | 1.001 (0.991–1.011) | 0.828 |
Stage (III vs. I or II) | 3.768 (2.235–6.354) | <0.001 | 3.693 (2.059–6.624) | <0.001 |
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Jo, J.H.; Park, S.B.; Park, S.; Lee, H.S.; Kim, C.; Jung, D.E.; Song, S.Y. Novel Gastric Cancer Stem Cell-Related Marker LINGO2 Is Associated with Cancer Cell Phenotype and Patient Outcome. Int. J. Mol. Sci. 2019, 20, 555. https://doi.org/10.3390/ijms20030555
Jo JH, Park SB, Park S, Lee HS, Kim C, Jung DE, Song SY. Novel Gastric Cancer Stem Cell-Related Marker LINGO2 Is Associated with Cancer Cell Phenotype and Patient Outcome. International Journal of Molecular Sciences. 2019; 20(3):555. https://doi.org/10.3390/ijms20030555
Chicago/Turabian StyleJo, Jung Hyun, Soo Been Park, Semi Park, Hee Seung Lee, Chanyang Kim, Dawoon E. Jung, and Si Young Song. 2019. "Novel Gastric Cancer Stem Cell-Related Marker LINGO2 Is Associated with Cancer Cell Phenotype and Patient Outcome" International Journal of Molecular Sciences 20, no. 3: 555. https://doi.org/10.3390/ijms20030555