Comparative Analysis of Blood and Bone Marrow for the Detection of Circulating and Disseminated Tumor Cells and Their Prognostic and Predictive Value in Esophageal Cancer Patients
Abstract
:1. Introduction
2. Methods
2.1. Patient Cohort
2.2. Liquid Biopsy Collection
2.3. Nested RT-PCR and Analysis
2.4. Statistical Analysis
3. Results
3.1. Characterization of the Analyzed Patient Cohort
3.2. Detection Rates of CK20 mRNA by RT-PCR
3.3. Correlation of CK20 mRNA Detection Levels in Blood and Bone Marrow and Patient’s Survival
3.4. Correlation between CK20 mRNA Detection in Blood and Bone Marrow and Survival of EC Patients in Dependence on UICC Stage
3.5. Correlation between CK20 mRNA Detection in Blood and Bone Marrow and Survival in EC Patients without Neo-Adjuvant Chemotherapy
3.6. Detection Rates of CK20 mRNA in Liquid Biopsies of EC Patients Associated with Clinical Parameters
3.7. Results of the Multivariate Analyses
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
References
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Cycle | Denaturation | Primer Hybridization 1 | Elongation | Cycles |
---|---|---|---|---|
Touch-down-cycles | ||||
1 | 40 s/94 °C | 2 min/70 °C | 1 | |
2 | 40 s/94 °C | 1 min 55 s/69 °C | 1 | |
3 | 40 s/94 °C | 1 min 50 s/68 °C (66 °C) | 1 | |
4 | 40 s/94 °C | 1 min 45 s/67 °C (65 °C) | 1 | |
5 | 40 s/94 °C | 1 min 40 s/66 °C (64 °C) | 1 | |
6 | 40 s/94 °C | 1 min 35 s/65 °C (63 °C) | 1 | |
7 | 40 s/94 °C | 1 min 30 s/64 °C (62 °C) | 1 | |
8 | 40 s/94 °C | 1 min 25 s/63 °C (61 °C) | 1 | |
9 | 40 s/94 °C | 1 min 20 s/62 °C (60 °C) | 1 | |
External and internal PCR | ||||
10–30 31 | 40 s/94 °C | 1 min/61 °C (59 °C) | 1 min 30 s/72 °C 15 min/72 °C | 21 1 |
n (%) | |
---|---|
All Sex | 77 (100) |
male | 64 (83.1) |
female | 13 (16.9) |
Tumor site | |
Squamous cell carcinoma | 37 (48.1) |
adenocarcinoma | 39 (50.6) |
Neuroendocrine tumor | 1 (1.3) |
Neo-adjuvant treatment | |
yes | 32 (41.6) |
no | 45 (58.4) |
Adjuvant treatment | |
yes | 2 (2.5) |
no | 53 (68.8) |
unknown | 22 (28.6) |
Variable | Detection of CK20 mRNA in Blood (n = 74) | Correlation χ2-Test (p-Value) | |||
---|---|---|---|---|---|
Total | Positive (%) | Negative (%) | |||
Gender | Female | 13 | 7 (53.85%) | 6 (46.15%) | 0.152 |
Male | 61 | 20 (32.79%) | 41 (67.21%) | ||
UICC-stage (n = 70) | I | 16 | 5 (31.25%) | 11 (68.75%) | 0.447 |
II | 26 | 9 (34.62%) | 17 (65.38%) | ||
III | 18 | 6 (33.33%) | 12 (66.67%) | ||
IV | 10 | 6 (60%) | 4 (40%) | ||
T-category (n = 71) | I | 18 | 5 (27.78%) | 13 (72.22%) | 0.439 |
II | 22 | 7 (31.82%) | 15 (68.18%) | ||
III | 30 | 14 (46.67%) | 16 (53.33%) | ||
IV | 1 | 0 | 1 (100%) | ||
N-category (n = 74) | 0 | 33 | 13 (39.39%) | 20 (60.61%) | 0.641 |
I | 41 | 14 (34.15%) | 27 (65.85%) | ||
M-category (n = 73) | 0 | 63 | 21 (33.33%) | 42 (66.67%) | 0.105 |
I | 10 | 6 (60%) | 4 (40%) | ||
Neo-adjuvant therapy | Yes | 31 | 12 (38.71%) | 19 (61.29%) | 0.736 |
No | 43 | 15 (34.88%) | 28 (65.12%) | ||
Relapse | Yes | 34 | 15 (44.12%) | 19 (55.88%) | 0.209 |
No | 40 | 12 (30%) | 28 (70%) | ||
Tumor type (histological) (n = 73) | Adenocarcinoma SCC | 38 | 10 (26.32%) | 28 (73.68%) | 0.084 |
35 | 16 (45.71%) | 19 (54.29%) |
Variable | Detection of CK20 mRNA in Bone Marrow (n = 61) | Correlation χ2-Test (p-Value) | |||
---|---|---|---|---|---|
Total | Positive (%) | Negative (%) | |||
Gender | Female | 9 | 2 (22.22%) | 7 (77.78%) | 0.53 |
Male | 52 | 17 (32.69%) | 35 (67.31%) | ||
UICC-stage (n = 70) | I | 12 | 3 (25%) | 9 (75%) | 0.49 |
II | 24 | 10 (41.67%) | 14 (58.33%) | ||
III | 15 | 3 (20%) | 12 (80%) | ||
IV | 8 | 3 (37.5%) | 5 (62.5%) | ||
T-category (n = 71) | I | 14 | 5 (35.71%) | 9 (64.29%) | 0.82 |
II | 19 | 7 (36.84%) | 12 (63.16%) | ||
III | 25 | 7 (28%) | 18 (72%) | ||
IV | 1 | 0 | 1 (100%) | ||
N-category (n = 61) | 0 | 27 | 8 (29.63%) | 19 (70.37%) | 0.82 |
I | 34 | 11 (32.35%) | 23 (67.65%) | ||
M-category (n = 73) | 0 | 53 | 16 (30.19%) | 37 (69.81%) | 0.68 |
I | 8 | 3 (37.5%) | 5 (62.5%) | ||
Neo-adjuvant therapy | Yes | 25 | 7 (28%) | 18 (72%) | 0.66 |
No | 36 | 12 (33.33%) | 24 (66.67%) | ||
Relapse | Yes | 28 | 8 (28.57%) | 20 (71.43%) | 0.69 |
No | 33 | 11 (33.33%) | 22 (66.67%) | ||
Tumor type (histological) (n = 73) | Adenocarcinoma SCC | 29 | 7 (24.14%) | 22 (75.86%) | 0.34 |
31 | 11 (35.48%) | 20 (64.52%) |
Variable | Detection of CK20 mRNA in Blood and/or Bone Marrow (n = 77) | Correlation χ2-Test (p-Value) | |||
---|---|---|---|---|---|
Total | Positive (%) | Negative (%) | |||
Gender | Female | 13 | 8 (61.54%) | 5 (38.46%) | 0.448 |
Male | 64 | 32 (50%) | 32 (50%) | ||
UICC-stage (n = 70) | I | 16 | 7 (43.75%) | 9 (56.25%) | 0.038 |
II | 28 | 18 (64.29%) | 10 (35.71%) | ||
III | 19 | 6 (31.58%) | 13 (68.52%) | ||
IV | 10 | 8 (80%) | 2 (20%) | ||
T-category (n = 71) | I | 18 | 9 (50%) | 9 (50%) | 0.598 |
II | 23 | 14 (60.87%) | 9 (39.13%) | ||
III | 32 | 16 (50%) | 16 (50%) | ||
IV | 1 | 0 | 1 (100%) | ||
N-category (n = 77) | 0 | 34 | 19 (55.88%) | 15 (44.12%) | 0.539 |
I | 43 | 21 (48.84%) | 22 (51.16%) | ||
M-category (n = 73) | 0 | 66 | 32 (48.48%) | 34 (51.52%) | 0.063 |
I | 10 | 8 (80%) | 2 (20%) | ||
Neo-adjuvant therapy | Yes | 32 | 17 (53.13%) | 15 (46.87%) | 0.862 |
No | 45 | 23 (51.11%) | 22 (48.89%) | ||
Relapse | Yes | 36 | 19 (52.78%) | 17 (47.22%) | 0.891 |
No | 41 | 21 (51.22%) | 20 (48.78%) | ||
Tumor type (histological) (n = 73) | Adenocarcinoma SCC | 39 | 16 (41%) | 23 (59%) | 0.065 |
37 | 23 (62.16%) | 14 (37.84%) |
Variables | p-Value (Univariate) | p-Value (Multivariate) | Hazard Quotient |
---|---|---|---|
Overall survival | |||
CK20 detection in BM | 0.029 | 0.006 | 2.529 |
N-category | <0.001 | 0.357 | 1.53 |
UICC stage-group | <0.001 | <0.001 | 3.128 |
Tumor-specific survival | |||
CK20 detection in BM | 0.048 | 0.013 | 2.37 |
N-category | <0.001 | 0.548 | 1.31 |
UICC stage-group | <0.001 | 0.002 | 2.873 |
Relapse free survival | |||
N-category | <0.001 | 0.258 | 1.646 |
UICC stage-group | <0.001 | 0.001 | 2.913 |
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Richter, F.; Baratay, A.; Röder, C.; Egberts, J.-H.; Kalthoff, H.; Becker, T.; Sebens, S. Comparative Analysis of Blood and Bone Marrow for the Detection of Circulating and Disseminated Tumor Cells and Their Prognostic and Predictive Value in Esophageal Cancer Patients. J. Clin. Med. 2020, 9, 2674. https://doi.org/10.3390/jcm9082674
Richter F, Baratay A, Röder C, Egberts J-H, Kalthoff H, Becker T, Sebens S. Comparative Analysis of Blood and Bone Marrow for the Detection of Circulating and Disseminated Tumor Cells and Their Prognostic and Predictive Value in Esophageal Cancer Patients. Journal of Clinical Medicine. 2020; 9(8):2674. https://doi.org/10.3390/jcm9082674
Chicago/Turabian StyleRichter, Florian, Annette Baratay, Christian Röder, Jan-Hendrik Egberts, Holger Kalthoff, Thomas Becker, and Susanne Sebens. 2020. "Comparative Analysis of Blood and Bone Marrow for the Detection of Circulating and Disseminated Tumor Cells and Their Prognostic and Predictive Value in Esophageal Cancer Patients" Journal of Clinical Medicine 9, no. 8: 2674. https://doi.org/10.3390/jcm9082674