Background
Methods
The cancer genome atlas (TCGA) database
Tissue microarray (TMA) construction and clinicopathological features
Immunohistochemistry (IHC)
Evaluation of PD-1 and PD-L1 expression TCs and TILs in FUSCC cohort
Microsatellite (MS) status and mismatch repair (MMR) status
Statistical analysis
Results
Clinical characteristics of patients with CRC in TCGA cohort and FUSCC cohort
TCGA Cohort (N = 356) | FUSCC Cohort (N = 276) | |||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Cases | PD-1 |
P
a
| PD-L1 |
P
a
| Cases | PD-1 |
P
a
| PD-L1 |
P
a
| |||||
No. (%) | Low | High | Low | High | No. (%) | Low | High | Low | High | |||||
Age (years) | ||||||||||||||
≤ 60 | 137 (38.5) | 68 (41.2) | 69 (36.1) | 0.325 | 21 (38.2) | 116 (38.5) | 0.960 | 170 (61.6) | 100 (58.8) | 70 (66.0) | 0.231 | 85 (61.6) | 85 (61.6) | 1.000 |
> 60 | 219 (35.7) | 97 (58.8) | 122 (63.9) | 34 (61.8) | 185 (61.5) | 106 (8.4) | 70 (41.7) | 36 (34.0) | 53 (38.4) | 53 (38.4) | ||||
Gender | ||||||||||||||
Male | 199 (55.9) | 91 (55.2) | 108 (56.5) | 0.792 | 33 (60.0) | 166 (55.1) | 0.505 | 166 (60.1) | 102 (60.0) | 64 (60.4) | 0.950 | 88 (63.8) | 78 (56.5) | 0.219 |
Female | 157 (44.1) | 74 (44.8) | 83 (43.5) | 22 (40.0) | 135 (44.9) | 110 (39.9) | 68 (40.0) | 42 (39.6) | 50 (36.2) | 60 (43.5) | ||||
Primary site | ||||||||||||||
Colon | 269 (75.6) | 115 (69.7) | 154 (80.6) |
0.017
| 46 (83.6) | 223 (74.1) | 0.130 | 129 (46.7) | 84 (49.4) | 45 (42.5) | 0.260 | 65 (47.1) | 64 (46.4) | 0.904 |
Rectum | 87 (24.4) | 50 (30.3) | 37 (19.4) | 9 (16.4) | 78 (25.9) | 147 (53.3) | 86 (50.6) | 61 (57.5) | 73 (52.9) | 74 (53.6) | ||||
Histological type | ||||||||||||||
Adenocarcinoma | 316 (88.8) | 149 (90.3) | 167 (87.4) | 0.393 | 50 (90.9) | 266 (88.4) | 0.584 | 261 (94.6) | 158 (92.9) | 103 (97.2) | 0.132 | 132 (95.6) | 129 (93.5) | 0.426 |
Mucinous/SRCC | 40 (11.2) | 16 (9.7) | 24 (12.6) | 5 (9.1) | 35 (11.6) | 15 (5.4) | 12 (7.1) | 3 (2.8) | 6 (4.3) | 9 (6.5) | ||||
T stage | ||||||||||||||
Tis-T2 | 68 (19.1) | 29 (17.6) | 39 (20.4) | 0.386 | 9 (16.4) | 78 (25.9) | 0.844 | 43 (15.6) | 24 (14.1) | 19 (17.9) | 0.588 | 19 (13.8) | 24 (17.4) |
0.044
|
T3 | 242 (68.0) | 118 (71.5) | 124 (64.9) | 39 (70.9) | 203 (67.4) | 32 (19.6) | 32 (18.8) | 22 (20.8) | 20 (14.5) | 34 (24.6) | ||||
T4 | 46 (12.9) | 18 (10.9) | 28 (14.7) | 7 (12.7) | 39 (13.0) | 179 (64.8) | 114 (67.1) | 65 (61.3) | 99 (71.7) | 80 (58.0) | ||||
N stage | ||||||||||||||
N0 | 194 (54.5) | 83 (50.3) | 111 (58.1) | 0.185 | 23 (41.8) | 171 (56.8) | 0.101 | 120 (43.5) | 70 (41.2) | 50 (47.2) | 0.432 | 57 (41.3) | 63 (45.7) | 0.767 |
N1 | 98 (27.5) | 53 (32.1) | 45 (23.6) | 18 (32.7) | 80 (26.6) | 83 (30.1) | 49 (28.8) | 33 (31.1) | 43 (31.2) | 40 (29.0) | ||||
N2 | 64 (18.0) | 29 (17.6) | 35 (18.3) | 14 (25.5) | 50 (16.6) | 73 (26.4) | 50 (29.4) | 23 (21.7) | 38 (27.5) | 35 (25.4) | ||||
M stage | ||||||||||||||
M0 | 245 (68.8) | 109 (66.1) | 136 (71.2) | 0.107 | 34 (61.8) | 211 (70.1) | 0.284 | 234(84.8) | 137 (80.6) | 97 (91.5) |
0.01
| 108(78.3) | 126 (91.3) |
0.003
|
M1 | 48 (13.5) | 29 (17.6) | 19 (9.9) | 11 (20.0) | 37 (12.3) | 42(15.2) | 33 (19.4) | 9 (8.5) | 30(21.7) | 12 (8.7) | ||||
Mx | 63 (17.7) | 27 (16.4) | 36 (18.8) | 10 (18.2) | 53 (17.6) | |||||||||
Pathological grading | ||||||||||||||
Well/moderate | NA | 204(73.9) | 67 (73.6) | 137 (74.1) | 0.498 | 102(73.9) | 102 (73.9) | 1.000 | ||||||
Poor/anaplastic | 56(20.3) | 18 (19.8) | 38 (20.5) | 28(20.3) | 28 (20.3) | |||||||||
Unknown | 16(5.8) | 6 (6.6) | 10 (5.4) | 8(5.8) | 8 (5.8) | |||||||||
Venous invasion | ||||||||||||||
Negative | 235 (66.0) | 105 (63.6) | 130 (68.1) | 0.557 | 28 (50.9) | 207 (68.8) |
0.026
| 184(66.7) | 107 (62.9) | 77 (72.6) | 0.104 | 85(61.6) | 99 (71.7) | 0.193 |
Positive | 70 (19.7) | 33 (20.0) | 37 (19.4) | 14 (25.5) | 56 (18.6) | 88(31.9) | 59 (34.7) | 29 (27.4) | 51(37.0) | 37 (26.8) | ||||
Unknown | 51 (14.3) | 27 (16.4) | 24 (12.6) | 13 (23.6) | 38 (12.6) | 4(1.4) | 4 (2.4) | 0 (0.0) | 2(1.4) | 2 (1.4) | ||||
Nervous invasion | ||||||||||||||
Negative | NA | 231 (83.7) | 75 (82.4) | 156 (84.3) | 0.181 | 118 (85.5) | 231 (83.7) | 0.337 | ||||||
Positive | 45 (16.3) | 16 (17.6) | 29 (15.7) | 20 (14.5) | 45 (16.3) | |||||||||
TILs extent | ||||||||||||||
Low | 47 (13.2) | 33 (20.0) | 14 (7.3) |
0.002
| 16 (29.1) | 31 (10.3) |
<0.001
| 129 (46.7) | 85 (50.0) | 44 (41.5) | 0.169 | 59 (42.8) | 70 (50.7) | 0.184 |
High | 93 (26.1) | 42 (25.5) | 51 (26.3) | 17 (30.9) | 76 (25.2) | 147 (53.3) | 85 (50.0) | 62 (58.5) | 79 (57.2) | 68 (49.3) | ||||
Unknown | 216 (60.7) | 90 (54.5) | 126 (66.0) | 22 (40.0) | 194 (64.5) | |||||||||
No. of LNs dissected | ||||||||||||||
< 12 | 80 (22.5) | 42 (25.5) | 38 (19.9) | 0.219 | 14 (25.5) | 56 (18.6) | 0.658 | 39 (14.1) | 24 (14.1) | 15 (14.2) | 0.994 | 16 (11.6) | 23 (16.7) | 0.226 |
≥ 12 | 247 (69.4) | 107(64.8) | 140 (73.3) | 38 (69.1) | 209 (69.4) | 237 (85.9) | 146 (85.9) | 91 (85.8) | 122 (88.4) | 115 (83.3) | ||||
Unknown | 29 (8.1) | 16 (9.7) | 13 (6.8) | 3 (5.5) | 26 (8.6) | |||||||||
MS status/MMR status | ||||||||||||||
MSS/MMR-proficient | 243 (68.3) | 123 (74.5) | 120 (62.8) |
0.018
| 39 (70.9) | 204 (67.8) | 0.646 | 176 (63.8) | 102 (60.0) | 74 (69.8) | 0.099 | 93 (67.4) | 83 (60.1) | 0.21 |
MSI/MMR-deficient | 113 (31.7) | 42 (25.5) | 71 (37.2) | 16 (29.1) | 97 (32.2) | 100 (36.2) | 68 (40.0) | 32 (30.2) | 45 (32.6) | 55 (39.9) | ||||
CEA (μl/ml) | ||||||||||||||
≤ 5 | 144 (40.4) | 59 (35.8) | 85 (44.5) | 0.151 | 13 (23.6) | 131 (43.5) |
0.007
| 170 (61.6) | 106 (62.4) | 64 (60.4) | 0.532 | 80 (58.0) | 90 (65.2) | 0.382 |
> 5 | 85 (23.9) | 39 (23.6) | 46 (24.1) | 13 (23.6) | 72 (23.9) | 95 (34.4) | 59 (34.7) | 36 (34.0) | 51 (37.0) | 44 (31.9) | ||||
Unknown | 127 (35.7) | 67 (40.6) | 60 (31.4) | 29 (52.7) | 98 (32.6) | 11 (4.0) | 5 (2.9) | 6 (5.7) | 7 (5.1) | 4 (2.9) | ||||
Adjuvant chemotherapy | ||||||||||||||
Yes | NA | 189 (68.5) | 58 (63.7) | 131 (70.8) | 0.082 | 89 (64.5) | 100 (72.5) | 0.05 | ||||||
No | 49 (17.8) | 15 (16.5) | 34 (18.4) | 23 (16.7) | 26 (18.8) | |||||||||
Unknown | 38 (13.7) | 18 (19.8) | 20 (10.8) | 26 (18.8) | 12 (8.7) |
Expression pattern of PD-1 and PD-L1
Prognostic significance of PD-1 and PD-L1 expression
Variablesa
| Univariate analysis HR (95 % CI) |
P
b
| Multivariate analysis HR (95 % CI) |
P
b
|
---|---|---|---|---|
Age (years) | ||||
≤ 60 | 1.000 | 0.202 | 1.000 | 0.937 |
> 60 | 1.366 (0.846–2.204) | 1.021 (0.606–1.720) | ||
Gender | ||||
Male | 1.000 | 0.132 | 1.000 | 0.205 |
Female | 0.687 (0.421–1.120) | 0.718 (0.431–1.198) | ||
Tumor location | ||||
Colon | 1.000 | 0.149 | ||
Rectum | 0.580 (0.277–1.215) | |||
Histological type | ||||
Adenocarcinoma | 1.000 | 0.148 | ||
Mucinous/SRCC | 1.611 (0.844–3.076) | |||
T stage | ||||
Tis-T2 | 1.000 |
<0.001
| 1.000 |
<0.001
|
T3 | 1.463 (0.618–3.465) | 0.832 (0.330–2.053) | ||
T4 | 8.584 (3.316–22.222) | 2.912 (1.242–8.093) | ||
N stage | ||||
N0 | 1.000 |
<0.001
| 1.000 | 0.583 |
N1 | 2.311 (1.306–4.091) | 1.430 (0.724–2.823) | ||
N2 | 3.098 (1.718–5.588) | 1.213 (0.557–2.642) | ||
M stage | ||||
M0 | 1.000 |
<0.001
| 1.000 | 0.115 |
M1 | 4.013 (2.252–7.152) | 2.062 (1.007–4.221) | ||
Mx | 1.996 (1.083–3.676) | 1.701 (0.837–3.458) | ||
Venous invasion | ||||
Negative | 1.000 |
0.006
| 1.000 | 0.145 |
Positive | 2.411 (1.400–4.151) | 2.008 (1.000–4.033) | ||
Unknown | 1.625 (0.842–3.137) | 1.362 (0.655–2.830) | ||
TILs extent | ||||
Low | 1.000 | 0.541 | ||
High | 0.746 (0.357–1.558) | |||
Unknown | 0.737 (0.422–1.286) | |||
No. of LNs dissected | ||||
< 12 | 1.000 | 0.145 | ||
≥ 12 | 2.311 (0.542–9.853) | |||
Unknown | 1.432 (0.347–5.917) | |||
MS status | ||||
MSS | 1.000 | |||
MSI | 1.159 (0.710–1.892) | 0.557 | ||
CEA (μl/ml) | ||||
≤ 5 | 1.000 | 1.000 |
0.011
| |
> 5 | 2.146 (0.978–4.708) |
0.003
| 1.339 (0.582–3.081) | |
Unknown | 3.086 (1.617–5.891) | 2.778 (1.337–5.772) | ||
PD-1 | ||||
Low | 1.000 | 1.000 | 0.68 | |
High | 0.591 (0.364–0.961) |
0.034
| 0.883 (0.488–1.596) | |
PD-L1 | ||||
Low | 1.000 | 1.000 | 0.148 | |
High | 0.463 (0.279–0.768) |
0.003
| 0.626 (0.332–1.181) |
Variablesa
| OS | DFS | ||||||
---|---|---|---|---|---|---|---|---|
Univariate analysis |
P
b
| Multivariate analysis |
P
b
| Univariate analysis |
P
b
| Multivariate analysis |
P
b
| |
HR (95 % CI) | HR (95 % CI) | HR (95 % CI) | HR (95 % CI) | |||||
Age (years) | ||||||||
≤ 60 | 1.000 | 0.367 | 1.000 | 0.564 | 1.000 | 0.593 | 1.000 | 0.633 |
> 60 | 1.224 (0.789–1.900) | 1.154 (0.709–1.879) | 1.115 (0.749–1.661) | 1.113 (0.717–1.730) | ||||
Genderb
| ||||||||
Male | 1.000 | 0.418 | 1.000 | 0.493 | 1.000 | 0.238 | 1.000 | 0.305 |
Female | 0.829 (0.526–1.306) | 0.843 (0.518–1.374) | 0.781 (0.518–1.178) | 0.793 (0.509–1.235) | ||||
Tumor location | ||||||||
Colon | 1.000 | 0.061 | 1.000 | 0.223 | ||||
Rectum | 0.658 (0.424–1.020) | 0.784 (0.529–1.160) | ||||||
Histological type | ||||||||
Adenocarcinoma | 1.000 | 0.781 | 1.000 | 0.765 | ||||
Mucinous/SRCC | 0.867 (0.317–2.369) | 0.872 (0.355–2.143) | ||||||
T stage | ||||||||
Tis-T2 | 1.000 |
<0.001
| 1.000 |
0.017
| 1.000 |
<0.001
| 1.000 |
0.032
|
T3 | 4.450 (0.975–20.312) | 6.329 (1.325–30.238) | 2.979 (0.971–9.138) | 3.471 (1.074–11.221) | ||||
T4 | 10.534 (2.581–42.992) | 8.734 (1.948–39.161) | 6.524 (2.390–17.805) | 4.282 (1.443–12.710) | ||||
N stage | ||||||||
N0 | 1.000 |
<0.001
| 1.000 | 0.269 | 1.000 |
<0.001
| 1.000 | 0.089 |
N1 | 2.770 (1.537–4.992) | 2.011 (0.982–4.117) | 2.927 (1.721–4.979) | 2.251 (1.202–4.215) | ||||
N2 | 4.146 (2.336–7.357) | 1.931 (0.903–4.130) | 4.212 (2.510–7.069) | 1.968 (0.984–3.934) | ||||
M stage | ||||||||
M0 | 1.000 |
<0.001
| 1.000 |
0.024
| 1.000 |
<0.001
| 1.000 |
<0.001
|
M1 | 9.094 (5.757–14.367) | 2.833 (1.137–6.083) | 10.198 (6.648–15.642) | 4.377 (1.999–9.585) | ||||
Pathological grading | ||||||||
Well/moderate | 1.000 | 0.378 | 1.000 | 0.729 | ||||
Poor/anaplastic | 1.278 (0.762–2.144) | 1.137 (0.706–1.832) | ||||||
Unknown | 0.576 (0.180–1.836) | 0.784 (0.317–1.941) | ||||||
Venous invasion | ||||||||
Negative | 1.000 |
<0.001
| 1.000 | 0.987 | 1.000 |
<0.001
| 1.000 | 0.55 |
Positive | 2.448 (1.578–3.796) | 0.986 (0.578–1.681) | 2.216 (1.492–3.291) | 0.760 (0.464–1.246) | ||||
Unknown | 1.043 (0.143–7.587) | 1.163 (0.154–8.756) | 0.813 (0.112–5.884) | 0.821 (0.110–6.136) | ||||
Nervous invasion | ||||||||
Negative | 1.000 | 0.059 | 1.000 |
0.008
| 1.000 | 0.199 | ||
Positive | 1.641 (0.981–2.743) | 1.186 (1.175–2.949) | 1.375 (0.846–2.237) | |||||
No. of LNs dissected | ||||||||
< 12 | 1.000 | 0.249 | 1.000 | 0.297 | ||||
≥ 12 | 0.719 (0.410–1.260) | 0.757 (0.449–1.276) | ||||||
MMR status | ||||||||
MMR-proficient | 1.000 | 0.726 | 1.000 | 0.792 | ||||
MMR-deficient | 1.084 (0.692–1.696) | 0.947 (0.632–1.418) | ||||||
CEA (μl/ml) | ||||||||
≤ 5 | 1.000 |
<0.001
| 1.000 |
0.041
| 1.000 |
<0.001
| 1.000 | 0.096 |
> 5 | 2.468 (1.572–3.875) | 1.386 (0.824–2.331) | 2.768 (1.845–4.152) | 1.529 (0.956–2.444) | ||||
Unknown | 2.850 (1.118–7.265) | 3.513 (1.292–9.558) | 2.472 (0.979–6.246) | 2.230 (0.848–5.863) | ||||
Adjuvant chemotherapy | ||||||||
Yes | 1.000 |
<0.001
| 1.000 |
0.019
| 1.000 |
<0.001
| 1.000 | 0.366 |
No | 0.670 (0.315–1.425) | 1.835 (0.735–4.584) | 0.568 (0.281–1.146) | 1.419 (0.609–3.307) | ||||
Unknown | 8.344 (5.142–13.542) | 3.130 (1.345–7.282) | 6.990 (4.495–10.870) | 1.645 (0.755–3.584) | ||||
TILs-PD-1 | ||||||||
Low | 1.000 |
0.007
| 1.000 |
0.006
| 1.000 |
0.003
| 1.000 |
0.015
|
High | 0.500 (0.302–0.828) | 0.480 (0.283–0.814) | 0.502 (0.319–0.789) | 0.559 (0.350–0.893) | ||||
TCs-PD-L1 | ||||||||
Low | 1.000 |
0.027
| 1.000 | 0.852 | 1.000 |
0.005
| 1.000 | 0.66 |
High | 0.606 (0.389–0.944) | 1.048 (0.639–1.719) | 0.558 (0.373–0.835) | 0.904 (0.577–1.417) |