Introduction
Material and methods
Patients
Expression analysis
Gene name | Primer and probe sequence | Primer and probe length | Product length |
---|---|---|---|
HPRT1
| F: AGCCTAAGATGAGAGTTC | 18 | 88 |
R: CACAGAACTAGAACATTGATA | 21 | ||
FAM-CATCTGGAGTCCTATTGACATCGC-TAMRA | 24 | ||
SOCS1
| F: TGGCCCCTTCTGTAGGATGG | 20 | 109 |
R: GGAGGAGGAAGAGGAGGAAGG | 21 | ||
FAM-TGGCCCCTTCTGTAGGATGG-TAMRA | 20 | ||
SOCS2
| F: ACGCGAACCCTTCTCTGACC | 20 | 99 |
R: CATTCCCGGAGGGCTCAAGG | 20 | ||
FAM-CTCGGGCGGCCACCTGTCTTTGC-TAMRA | 23 | ||
SOCS3
| F: GTGGAGAGGCTGAGGGACTC | 20 | 111 |
R: GGCTGACATTCCCAGTGCTC | 20 | ||
FAM-CACCAAGCCAGCCCACAGCCAGG-TAMRA | 23 | ||
SOCS5
| F: GTGACTCGGAAGAGGATACAACC | 23 | 91 |
R: CTAACATGGGTATGGCTGTCTCC | 23 | ||
FAM-CGCTGCTTCTGCCTCCGTGACTGC-TAMRA | 24 |
Estrogen receptor (ER)/progesterone receptor (PR)
HER2/neu
Ki-67
Statistical analysis
Results
General demographic data of patients
Variables | Values |
---|---|
Age (years) (mean ± SD) | 51.79 ± 13.54 (29–81) |
Menarche age (years) (mean ± SD) | 13 ± 1.65 (10–18) |
Menopause age (years) (mean ± SD) | 44.91 ± 14.91 (38–60) |
First pregnancy age (years) (mean ± SD) | 18.04 ± 8.36 (14–32) |
Breast feeding duration (months) (mean ± SD) | 41.62 ± 34.1 (3–120) |
Cancer stage (%) | |
I | 30.8 |
II | 28.8 |
III | 30.8 |
IV | 9.6 |
Overall grade (%) | |
I | 17 |
II | 49 |
III | 34 |
Mitotic rate (%) | |
I | 45.2 |
II | 42.9 |
III | 11.9 |
Tumor size (%) | |
< 2 cm | 32 |
≥ 2 cm, < 5 cm | 66 |
≥ 5 cm | 2 |
Estrogen receptor (%) | |
Positive | 87.8 |
Negative | 12.2 |
Progesterone receptor (%) | |
Positive | 77.1 |
Negative | 22.9 |
Her2/neu expression (%) | |
Positive | 25 |
Negative | 75 |
Ki67 expression (%) | |
Positive | 100 |
Negative | 0 |
Relative expression of SOCS in tumoral tissues vs. ANCTs
Association between relative expression of genes and patients’ clinicopathological data
SOCS1 upregulation | SOCS1 downregulation | P value | SOCS2 upregulation | SOCS2 downregulation | P value | SOCS3 upregulation | SOCS3 downregulation | P value | SOCS5 upregulation | SOCS5 downregulation | P value | |
---|---|---|---|---|---|---|---|---|---|---|---|---|
Age | 0.35 | 0.5 | 0.84 | 0.68 | ||||||||
< 55 years | 11 (32.4%) | 23 (67.6%) | 9 (26.5%) | 25 (73.5%) | 11 (32.4%) | 23 (67.6%) | 10 (29.4%) | 24 (70.6%) | ||||
≥ 55 years | 9 (45%) | 11 (55%) | 7 (35%) | 13 (65%) | 7 (35%) | 13 (65%) | 5 (25%) | 15 (75%) | ||||
Stage | 0.95 | 0.81 | 0.8 | 0.86 | ||||||||
1 | 5 (31.3%) | 11 (68.7%) | 5 (31.3%) | 11 (68.7%) | 4 (25%) | 12 (75%) | 3 (18.8%) | 13 (81.2%) | ||||
2 | 6 (40%) | 9 (60%) | 6 (40%) | 9 (60%) | 6 (40%) | 9 (60%) | 5 (33.3%) | 10 (66.7%) | ||||
3 | 7 (43.8%) | 9 (56.3%) | 4 (25%) | 12 (75%) | 5 (31.2%) | 11 (68.8%) | 5 (31.2%) | 11 (68.8%) | ||||
4 | 2 (40%) | 3 (60%) | 1 (20%) | 4 (80%) | 2 (40%) | 3 (60%) | 1 (20%) | 4 (80%) | ||||
Histological grade | 0.32 | 0.18 | 0.08 | 0.19 | ||||||||
1 | 2 (25%) | 6 (75%) | 1 (12.5%) | 7 (87.5%) | 0 (0%) | 8 (100%) | 0 (0%) | 8 (100%) | ||||
2 | 12 (52.2%) | 11 (47.8%) | 10 (43.5%) | 13 (56.5%) | 9 (39.1%) | 14 (60.9%) | 8 (34.8%) | 15 (65.2%) | ||||
3 | 5 (31.3%) | 11 (68.8%) | 3 (18.8%) | 13 (81.2%) | 7 (43.8%) | 9 (56.2%) | 4 (25%) | 12 (75%) | ||||
Mitotic rate | 0.36 | 0.03 | 0.9 | 0.16 | ||||||||
1 | 10 (52.6%) | 9 (47.4%) | 10 (52.6%) | 9 (47.4%) | 8 (42.1%) | 11 (57.9%) | 8 (42.1%) | 11 (57.9%) | ||||
2 | 6 (33.3%) | 12 (66.7%) | 4 (22.2%) | 14 (77.8%) | 6 (33.3%) | 12 (66.7%) | 4 (22.2%) | 14 (77.8%) | ||||
3 | 1 (20%) | 4 (80%) | 0 (0%) | 5 (100%) | 2 (40%) | 3 (60%) | 0 (0%) | 5 (100%) | ||||
Tumor size | 0.26 | 0.29 | 1 | 0.38 | ||||||||
< 2 | 4 (25%) | 12 (75%) | 5 (31.3%) | 11 (68.8%) | 5 (31.3%) | 11 (68.8%) | 4 (25%) | 12 (75%) | ||||
2–5 | 15 (45.5%) | 18 (54.5%) | 9 (27.3%) | 24 (72.7%) | 11 (33.3%) | 12 (66.7%) | 8 (24.2%) | 25 (75.8%) | ||||
> 5 | 0 (0%) | 1 (100%) | 1 (100%) | 0 (0%) | 0 (0%) | 1 (100%) | 1 (100%) | 0 (0%) | ||||
ER state | 0.81 | 0.63 | 1 | 0.36 | ||||||||
Positive | 16 (38.1%) | 26 (61.9%) | 13 (31%) | 29 (69%) | 14 (33.3%) | 28 (66.7%) | 10 (23.8%) | 32 (76.2%) | ||||
Negative | 3 (42.9%) | 4 (57.1%) | 2 (28.6%) | 5 (71.4%) | 2 (28.6%) | 5 (71.4%) | 3 (42.9%) | 4 (57.1%) | ||||
PR state | 0.8 | 0.72 | 0.33 | 0.45 | ||||||||
Positive | 15 (40.5%) | 22 (59.5%) | 11 (29.7%) | 26 (70.3%) | 11 (29.7%) | 26 (70.3%) | 9 (24.3%) | 28 (75%) | ||||
Negative | 4 (36.4%) | 7 (63.6%) | 4 (36.4%) | 7 (63.6%) | 5 (45.5%) | 6 (54.5%) | 4 (36.4%) | 7 (63.6%) | ||||
Her2 state | 0.39 | 0.85 | 0.15 | 0.18 | ||||||||
Positive | 6 (50%) | 6 (50%) | 4 (33.3%) | 8 (66.7%) | 6 (50%) | 6 (50%) | 5 (41.7%) | 7 (58.3%) | ||||
Negative | 13 (36.1%) | 23 (63.9%) | 11 (30.6%) | 25 (69.4%) | 10 (27.8%) | 26 (72.2%) | 8 (22.2%) | 28 (77.8%) | ||||
Breast feeding duration (months) | 0.3 | 0.84 | 0.08 | 0.19 | ||||||||
0 | 4 (50%) | 4 (50%) | 2 (25%) | 6 (75%) | 5 (62.5%) | 3 (37.5%) | 4 (50%) | 4 (50%) | ||||
1–30 | 8 (53.3%) | 7 (46.7%) | 6 (40%) | 9 (60%) | 6 (40%) | 9 (60%) | 6 (40%) | 9 (60%) | ||||
31–60 | 5 (27.8%) | 13 (72.2%) | 5 (27.8%) | 13 (72.2%) | 6 (33.3%) | 12 (66.7%) | 3 (16.7%) | 15 (83.3%) | ||||
61–120 | 3 (25%) | 9 (75%) | 3 (25%) | 9 (75%) | 1 (8.3%) | 11 (91.7%) | 2 (16.7%) | 10 (83.3%) |
SOCS1 expression (mean (SD)) | P value | SOCS2 expression (mean (SD)) | P value | SOCS3 expression (mean (SD)) | P value | SOCS5 expression (mean (SD)) | P value | |
---|---|---|---|---|---|---|---|---|
Age | ||||||||
< 55 years vs. ≥ 55 years | 496.1 (2.6) vs. 592.9 (2.6) | 0.88 | 1.5 (7.9) vs. 10.9 (17.2) | 0.43 | 199.5 (542.6) vs. 84.7 (372.8) | 0.4 | 16.7 (44.9) vs. 1.1 (4.9) | 0.33 |
ER status | ||||||||
ER (+) vs. ER (−) | 675.4 (2.9) vs. 0.6 (0.8) | 0.55 | 1.2 (7.1) vs. 5.1 (7.7) | 0.64 | 192.5 (545.1) vs. 55.1 (127.2) | 0.51 | 537.3 (3.3) vs. 1.5 (1.5) | 0.68 |
PR status | ||||||||
PR (+) vs. PR (−) | 766.1 (3.1) vs. 2.5 (6.9) | 0.42 | 1.4 (7.5) vs. 4 (5.9) | 0.53 | 216.6 (577.2) vs. 41.5 (102.1) | 0.08 | 609.9 (3.6) vs. 1.2 (1.3) | 0.58 |
HER2 status | ||||||||
HER2 (+) vs. HER2 (−) | 20 (25.1) vs. 781.5 (3.1) | 0.41 | 10.3 (17.8) vs. 1.4 (7.6) | 0.51 | 223.3 (763) vs. 160.9 (411.2) | 0.71 | 3.4 (5.6) vs. 626 (3.6) | 0.56 |
Tumor grade | ||||||||
Grade 1 vs. 2 | 3.4 (6.2) vs. 13.7 (23.4) | 0.004 | 6.5 (1.6) vs. 13.8 (20.6) | 0.04 | 596 (735) vs. 25.7 (63.9) | 0.01 | 2.8 (7.7) vs. 3.8 (7.1) | 0.08 |
Grade 1 vs. 3 | 3.4 (6.2) vs. 8.1 (19.2) | 0.007 | 6.5 (1.6) vs. 10.8 (17.6) | 0.05 | 596 (735) vs. 194.5 (659.3) | 0.1 | 2.8 (7.7) vs. 2.3 (3.9) | 0.1 |
Grade 2 vs. 3 | 13.7 (23.4) vs. 8.1 (19.2) | 1 | 13.8 (20.6) vs. 10.8 (17.6) | 1 | 25.7 (63.9) vs. 194.5 (659.3) | 0.5 | 3.8 (7.1) vs. 2.3 (3.9) | 1 |
Correlation between relative expressions of genes
SOCS5
|
SOCS3
|
SOCS2
| ||
---|---|---|---|---|
SOCS1
| Tumor | 0.59** | 0.65** | 0.81** |
ANCT | 0.96** | 0.85** | 0.52** | |
SOCS2
| Tumor | 0.02 | 0.5** | |
ANCT | 0.69** | 0.88** | ||
SOCS3
| Tumor | 0.43* | ||
ANCT | 0.95** |
Partial correlation between expression of genes and patients’ age and tumor grade
SOCS1
|
SOCS2
|
SOCS3
|
SOCS5
| |||||
---|---|---|---|---|---|---|---|---|
R
| P value |
R
| P value |
R
| P value |
R
| P value | |
Age (controlled for grade) | − 0.02 | 0.88 | − 0.03 | 0.84 | − 0.06 | 0.66 | − 0.007 | 0.96 |
Grade (controlled for age) | − 0.36 | 0.007 | − 0.28 | 0.02 | − 0.18 | 0.1 | − 0.25 | 0.04 |
ROC curve analysis
Estimate criterion | AUC |
J
a
| Sensitivity | Specificity | P valueb | |
---|---|---|---|---|---|---|
SOCS1
| > 2.7 | 0.6 | 0.2 | 40.7 | 79.6 | 0.05 |
SOCS2
| > − 2.4 | 0.64 | 0.25 | 88.9 | 37 | 0.007 |
SOCS3
| > − 0.9 | 0.63 | 0.24 | 72.2 | 51.9 | 0.01 |
SOCS5
| > − 0.8 | 0.67 | 0.25 | 75.9 | 50 | 0.0009 |
Combination of all genes | ≤ 0.5 | 0.68 | 0.35 | 64.8 | 70.4 | 0.0004 |