Background
Methods
Patients
Tissue microarray construction
Immunohistochemistry and staining evaluation
Statistical analysis
Results
MMR immunophenotype
n (%) | MLH1 | PMS2 | MSH2 | MSH6 |
---|---|---|---|---|
All | ||||
1 (5.0) | Intact | Loss | Intact | Uninterpretablea |
1 (5.0) | Loss | Loss | Intact | Loss |
3 (15.0) | Loss | Loss | Intact | Intact |
4 (20.0) | Intact | Loss | Intact | Intact |
3 (15.0) | Intact | Intact | Loss | Loss |
8 (40.0) | Intact | Intact | Intact | Loss |
I-type | ||||
1 (7.7) | Loss | Loss | Intact | Loss |
3 (23.1) | Loss | Loss | Intact | Intact |
4 (30.1) | Intact | Loss | Intact | Intact |
2 (15.4) | Intact | Intact | Loss | Loss |
3 (23.1) | Intact | Intact | Intact | Loss |
PB-type | ||||
1 (14.3) | Intact | Loss | Intact | Uninterpretablea |
1 (14.3) | Intact | Intact | Loss | Loss |
5 (71.4) | Intact | Intact | Intact | Loss |
Associations of MMR immunophenotype with clinicopathological factors and tumor-infiltrating immune cells
Intestinal type | Pancreatobiliary type | All | |||||||
---|---|---|---|---|---|---|---|---|---|
pMMR (n = 50) | dMMR (n = 13) |
p
| pMMR (n = 102) | dMMR (n = 7) |
p
| pMMR (n = 152) | dMMR (n = 20) |
p
| |
Age | |||||||||
(Median, range) | 66.5 (38.0–79.0) | 67.0 (48.0–83.0) | 0.905 | 67.0 (44.0–81.0) | 62.0 (58.0–76.0) | 0.669 | 67.0 (38.0–81.0) | 65.5 (48.0–83.0) | 0.635 |
Sex | |||||||||
Women | 26 (52.0) | 8 (61.5) | 0.539 | 46 (45.1) | 4 (57.1) | 0.538 | 72 (47.4) | 12 (60.0) | 0.289 |
Men | 24 (48.0) | 5 (38.5) | 56 (54.9) | 3 (42.9) | 80 (52.6) | 8 (40.0) | |||
Tumor origin | |||||||||
Duodenum | 8 (16.0) | 6 (46.2) | 0.005 | 8 (5.3) | 6 (28.6) | 0.003 | |||
Ampulla intestinal type | 42 (84.0) | 7 (53.8) | 42 (27.6) | 7 (35.0) | |||||
Ampulla pancreatobiliary type | 17 (16.7) | 2 (28.6) | 0.860 | 17 (11.2) | 2 (10.0) | ||||
Distal bile duct | 44 (43.1) | 1 (14.3) | 44 (28.9) | 1 (5.0) | |||||
Pancreas | 41 (40.2) | 4 (57.1) | 41 (27.0) | 4 (20.0) | |||||
Tumor size mm | |||||||||
(Median, range) | 25.0 (5.0–60.0) | 40.0 (13.0–90.0) | 0.006 | 30.0 (9.0–70.0) | 30.0 (5.0–35.0) | 0.283 | 30.0 (5.0–70.0) | 33.0 (5.0–90.0) | 0.107 |
Differentiation grade | |||||||||
Well-moderate | 24 (48.0) | 7 (53.8) | 0.707 | 37 (36.3) | 4 (57.1) | 0.272 | 61 (40.1) | 11 (55.0) | 0.206 |
Poor | 26 (52.0) | 6 (46.2) | 65 (63.7) | 3 (42.9) | 91 (59.9) | 9 (45.0) | |||
T-stage | |||||||||
T1 | 4 (8.0) | 0 | 0.736 | 2 (2.0) | 0 | 0.966 | 6 (3.9) | 0 | 0.420 |
T2 | 9 (18.0) | 2 (15.4) | 11 (10.8) | 1 (14.3) | 20 (13.2) | 3 (15.0) | |||
T3 | 19 (38.0) | 6 (46.2) | 73 (71.6) | 5 (71.4) | 92 (60.5) | 11 (55.0) | |||
T4 | 18 (36.0) | 5 (38.5) | 16 (15.7) | 1 (14.3) | 34 (22.4) | 6 (30.0) | |||
N-stage | |||||||||
N0 | 24 (48.0) | 9 (69.2) | 0.348 | 26 (25.5) | 6 (85.7) | 0.002 | 50 (32.9) | 15 (75.0) | 0.002 |
N1 | 17 (34.0) | 2 (15.4) | 44 (43.1) | 1 (14.3) | 61 (40.1) | 3 (15.0) | |||
N2 | 9 (18.0) | 2 (15.4) | 32 (31.4) | 0 | 41 (27.0) | 2 (10.0) | |||
Margins | |||||||||
R0 | 13 (26.0) | 4 (30.8) | 0.730 | 6 (5.9) | 0 | 0.511 | 19 (12.5) | 4 (20.0) | 0.356 |
R1–Rx | 37 (74.0) | 9 (69.2) | 96 (94.1) | 7 (100.0) | 133 (87.5) | 16 (80.0) | |||
Perineural growth | |||||||||
No | 33 (66.0) | 11 (84.6) | 0.193 | 21 (20.6) | 3 (42.9) | 0.171 | 54 (35.5) | 14 (70.0) | 0.003 |
Yes | 17 (34.0) | 2 (15.4) | 81 (79.4) | 4 (57.1) | 98 (64.5) | 6 (30.0) | |||
Invasion of lymphatic vessels | |||||||||
No | 18 (36.0) | 11 (84.6) | 0.002 | 29 (28.4) | 5 (71.4) | 0.018 | 47 (30.9) | 16 (80.0) | 0.000 |
Yes | 32 (64.0) | 2 (15.4) | 72 (71.6) | 2 (28.6) | 105 (69.1) | 4 (20.0) | |||
Invasion of blood vessels | |||||||||
No | 45 (90.0) | 13 (100.0) | 0.235 | 67 (65.7) | 5 (71.4) | 0.757 | 112 (73.7) | 18 (90.0) | 0.111 |
Yes | 5 (10.0) | 0 | 35 (3437) | 2 (28.6) | 40 (26.3) | 2 (10.0) | |||
Growth in peripancreatic fat | |||||||||
No | 31 (62.0) | 10 (76.9) | 0.315 | 20 (19.6) | 4 (57.1) | 0.021 | 51 (33.6) | 14 (70.0) | 0.002 |
Yes | 19 (38.0) | 3 (23.1) | 82 (80.4) | 3 (42.9) | 101 (66.4) | 6 (30.0) | |||
Adjuvant chemotherapy | |||||||||
None | 35 (70.0) | 10 (76.9) | 0.094 | 46 (45.1) | 4 (57.1) | 0.177 | 81 (53.3) | 14 (70.0) | 0.040 |
5FU-analogue | 5 (10.0) | 0 | 8 (7.8) | 0 | 13 (8.6) | 0 (0.0) | |||
Gemcitabine | 7 (14.0) | 0 | 43 (42.2) | 2 (28.6) | 50 (32.9) | 2 (10.0) | |||
Gemcitabine + capecitabine | 0 | 1 (7.7) | 3 (2.9) | 0 | 3 (2.0) | 1 (5.0) | |||
Oxaliplatin + 5-FU analogue | 2 (4.0) | 2 (15.4) | 1 (1.0) | 0 | 3 (2.0) | 2 (10.0) | |||
Gemcitabine + oxaliplatin | 1 (2.0) | 0 | 1 (1.0) | 1 (12.5) | 2 (1.3) | 1 (5.0) | |||
Immune cells | |||||||||
CD3+ | 206 (6–795) | 246 (59–559) | 0.156 | 129 (22–546) | 44 (26–695) | 0.092 | 148.5 (6–795) | 227 (26–695) | 0.471 |
Missing | 0 | 0 | 0 | 0 | 0 | 0 | |||
CD8+ | 55 (2–180) | 114 (2–180) | 0.012 | 45 (4–175) | 37 (1–200) | 0.844 | 47.5 (2–180) | 101.5 (1–200) | 0.035 |
Missing | 1 | 0 | 3 | 0 | 4 | 0 | |||
FOXP3+ | 33 (0–110) | 40 (1–103) | 0.497 | 26 (1–137) | 15 (0–119) | 0.420 | 27.5 (0–137) | 38 (0–119) | 0.560 |
Missing | 0 | 0 | 2 | 1 | 2 | 1 | |||
CD68+ | 87 (29–350) | 74 (19–182) | 0.490 | 98 (25–230) | 97 (36–230) | 0.645 | 93 (25–350) | 82 (19–229) | 0.455 |
Missing | 0 | 0 | 1 | 1 | 1 | 1 | |||
CD163+ | 130 (35–250) | 120 (44–190) | 0.755 | 140 (49–275) | 162 (92–200) | 0.241 | 136 (35–275) | 131 (44–200) | 0.885 |
Missing | 1 | 0 | 4 | 1 | 5 | 1 | |||
CD56+ | 1 (0–9) | 2 (0–23) | 0.370 | 1 (0–12) | 3 (0–33) | 0.046 | 1 (0–12) | 2 (0–33) | 0.029 |
Missing | 0 | 0 | 0 | 0 | 0 | 0 |
Prognostic and potential predictive value of MMR immunophenotype
Intestinal type | Pancreatobiliary type | All | |||||||
---|---|---|---|---|---|---|---|---|---|
n (events) | Unadjusted | Adjusted | n (events) | Unadjusted | Adjusted | n (events) | Unadjusted | Adjusted | |
HR (95% CI) | HR (95% CI) | HR (95% CI) | HR (95% CI) | HR (95% CI) | HR (95% CI) | ||||
Age | |||||||||
Continuous | 63 (39) | 1.01 (0.98–1.05) | – | 109 (98) | 0.99 (0.97–1.02) | – | 172 (137) | 1.01 (0.99–1.03) | – |
Gender | |||||||||
Female | 35 (17) | 1.00 | 1.00 | 51 (45) | 1.00 | – | 86 (62) | 1.00 | – |
34 (16)* | |||||||||
Male | 28 (22) |
2.01 (1.05–3.84)
| 1.60 (0.79–3.24) | 58 (53) | 1.15 (0.77–1.71) | 86 (75) |
1.40 (1.00–1.97)
| ||
27 (21)* | |||||||||
Tumor origin | |||||||||
Intestinal | – | – | – | – | 63 (39) | 1.00 | 1.00 | ||
61 (37)* | |||||||||
Pancreatobiliary type | – | – | – | – | 109 (98) |
2.54 (1.73–3.73)
|
1.61 (1.01–2.55)
| ||
108 (98)* | |||||||||
Tumor size | |||||||||
Continuous | 63 (39) | 1.00 (0.98–1.02) | – | 109 (98) |
1.04 (1.02–1.06)
|
1.02 (1.00–1.04)
| 172 (137) |
1.02 (1.00–1.03)
| 1.01 (0.99–2.55) |
108 (98)* | 169 (135)* | ||||||||
T-stage | |||||||||
T1–T2 | 16 (8) | 1.00 | – | 15 (10) | 1.00 | 1.00 | 31 (18) | 1.00 | 1.00 |
14 (10)* | 28 (16)* | ||||||||
T3–T4 | 47 (31) | 1.72 (0.79–3.74) | 94 (88) |
2.47 (1.28–4.77)
| 1.06 (0.50–2.27) | 141 (119) |
2.34 (1.42–3.86)
| 1.21 (0.66–2.21) | |
94 (88)* | 141 (119)* | ||||||||
N-stage | |||||||||
N0 | 35 (19) | 1.00 | – | 32 (25) | 1.00 | 1.00 | 67 (44) | 1.00 | 1.00 |
31 (25)* | 64 (42)* | ||||||||
N1–2 | 28 (20) | 1.56 (0.83–2.94) | 77 (73) |
2.23 (1.41–3.54)
|
2.06 (1.16–3.65)
| 105 (93) |
2.19 (1.52–3.16)
| 1.34 (0.88–2.05) | |
77 (73)* | 105 (93)* | ||||||||
Differentiation grade | |||||||||
Well-moderate | 31 (17) | 1.00 | – | 42 (32) | 1.00 | 1.00 | 73 (49) | 1.00 | 1.00 |
41 (32)* | 71 (48)* | ||||||||
Poor | 32 (22) | 1.70 (0.90–3.21) | 67 (66) |
2.63 (1.70–4.09)
|
2.05 (1.24–3.39)
| 99 (88) |
2.24 (1.57–3.18)
|
1.65 (1.09–2.48)
| |
67 (66)* | 98 (87)* | ||||||||
Involved margins, status | |||||||||
R0 | 18 (5) | 1.00 | 1.00 | 7 (5) | 1.00 | – | 25 (10) | 1.00 | 1.00 |
17 (4)* | 23 (9)* | ||||||||
R1 and Rx | 45 (34) |
3.30 (1.29–8.46)
| 2.89 (0.94–8.85) | 102 (93) | 2.42 (0.98–6.00) | 147 (127) |
3.57 (1.87–6.83)
|
2.23 (1.10–4.51)
| |
44 (33)* | 146 (126)* | ||||||||
Lymphatic growth | |||||||||
Absent | 28 (10) | 1.00 | 1.00 | 35 (27) | 1.00 | 1.00 | 63 (37) | 1.00 | 1.00 |
28 (10)* | 34 (27)* | 62 (37)* | |||||||
Present | 35 (29) |
3.94 (1.90–8.18)
| 1.77 (0.77–4.09) | 74 (71) |
1.78 (1.13–2.79)
| 1.09 (0.65–1.83) | 109 (100) |
2.54 (1.73–3.73)
| 1.33 (0.85–2.07) |
33 (27)* | 74 (71)* | 107 (98)* | |||||||
Vascular growth | |||||||||
Absent | 58 (34) | 1.00 | 1.00 | 73 (62) | 1.00 | 1.00 | 131 (96) | 1.00 | 1.00 |
56 (32)* | 72 (62)* | 128 (94)* | |||||||
Present | 5 (5) |
6.68 (2.40–18.62)
|
4.69 (1.48–14.84)
| 36 (36) |
2.47 (1.61–3.80)
|
2.20 (1.38–3.52)
| 41 (41) |
3.49 (2.37–5.15)
|
2.18 (1.43–3.34)
|
5 (5)* | 36 (36)* | 41 (41)* | |||||||
Perineural growth | |||||||||
Absent | 43 (21) | 1.00 | 1.00 | 25 (19) | 1.00 | 1.00 | 68 (40) | 1.00 | 1.00 |
42 (20)* | 24 (19)* | 66 (39)* | |||||||
Present | 20 (18) |
2.81 (1.48–5.35)
| 1.36 (0.57–3.21) | 84 (79) |
1.96 (1.18–3.25)
| 0.88 (0.48–1.61) | 104 (97) |
2.96 (2.02–4.43)
| 1.06 (0.66–1.71) |
19 (17)* | 84 (79)* | 103 (96)* | |||||||
Growth in peripancreatic fat | |||||||||
Absent | 42 (20) | 1.00 | 1.00 | 25 (20) | 1.00 | 1.00 | 67 (40) | 1.00 | 1.00 |
40 (18)* | 24 (20)* | 64 (38)* | |||||||
Present | 21 (19) |
3.65 (1.86–7.16)
| 1.78 (0.73–4.33) | 84 (78) |
1.75 (1.06–2.88)
| 0.99 (0.56–1.77) | 105 (97) |
3.00 (2.03–4.43)
| 1.37 (0.85–2.22) |
21 (19)* | 84 (78)* | 105 (97)* | |||||||
Adjuvant treatment | |||||||||
None | 45 (30) | 1.00 | – | 50 (44) | 1.00 | – | 95 (74) | 1.00 | – |
Any | 18 (9) | 0.69 (0.33–1.47) | 59 (54) | 0.96 (0.64–1.42) | 77 (63) | 1.09 (0.78–1–53) | |||
MMR | |||||||||
pMMR | 49 (34) | 1.00 | 1.00 | 101 (93) | 1.00 | 1.00 | 150 (127) | 1.00 | 1.00 |
49 (34)* | 101 (93)* | 150 (127)* | |||||||
dMMR | 12 (3) |
0.20 (0.06–0.68)
| 0.36 (0.09–1.42) | 7 (5) | 0.57 (0.23–1.42) | 1.24 (0.46–3.37) | 19 (8) |
0.28 (0.13–0.57)
| 0.52 (0.23–1.19) |
12 (3)* | 7 (5)* | 19 (8)* |