Background
Methods
Patients
Pathological review
Histological definition of micropapillary components
Tumor tissue microarray (TMA) synthesis
Immunohistochemistry
Classification/Antibody | Clone | Dilution | Source |
---|---|---|---|
Cellular adhesion molecules
| |||
E-cadherin | NCH-38 | 1:100 | DakoCytomation, Carpinteria, CA, USA |
CD44 | DF1485 | 1:400 | Novocastra, Newcastle upon Tyne, UK |
Laminin5γ2 | 4G1 | 1:50 | DakoCytomation, Glostrup, Denmark |
Growth factor | |||
VEGF-C | Polyclonal | 1:50 | Abcam, Cambridge, UK |
Apoptosis-associated proteins | |||
bcl2 | 124 | 1:50 | DakoCytomation, Glostrup, Denmark |
p53 | DO-7 | Pre-diluted | Nichirei, Tokyo, Japan |
cleaved caspase-3 | Polyclonal | 1:400 | Cell signaling, Danvers, MA, USA |
Mucin-related proteins | |||
MUC1 | Ma695 | 1:100 | Novocastra, Newcastle upon Tyne, UK |
MUC6 | CLH5 | 1:100 | Novocastra, Newcastle upon Tyne, UK |
Hypoxia induced protein | |||
HIF-1α | EP1215Y | 1:500 | Abcam, Cambridge, UK |
Others | |||
TTF-1 | 8G7G3/1 | 1:100 | DakoCytomation, Carpinteria, CA, USA |
SP-A | PE10 | 1:100 | Dako, Kyoto, Japan |
Vimentin | V9 | Pre-diluted | DakoCytomation, Carpinteria, CA, USA |
Ki-67 | MIB-1 | 1:50 | Dako, Glostrup, Denmark |
LYVE1 | 15A5B2 | 1:400 | Oriental Yeast, Tokyo, Japan |
c-Met | EP1454Y | 1:200 | Abcam, Cambridge, UK |
Phospho-c-Met | Polyclonal | 1:800 | Stressgen, Ann Arbor, MI, USA |
Calculation of staining scores
Mutation analysis
Statistical analysis
Results
Clinicopathological characteristics of patients with SMPC
all | % | SMPC | P value | ||||
---|---|---|---|---|---|---|---|
(-) | % | (+) | % | ||||
No. | 559 | 540 | 97 | 19 | 3 | ||
Age | |||||||
Median | 67 | 67 | 67 | 0.219* | |||
Range | 23-87 | 23-87 | 40-76 | ||||
Gender | |||||||
Female | 288 | 52 | 282 | 52 | 6 | 32 | 0.077** |
Male | 271 | 48 | 258 | 48 | 13 | 68 | |
Smoking status | |||||||
Nonsmoker | 284 | 51 | 276 | 51 | 8 | 42 | 0.596** |
Smoker | 275 | 49 | 264 | 49 | 11 | 58 | |
BI Average | 369 | 364 | 502 | ||||
Tumor size | |||||||
Average(mm) | 25 | 25 | 35 | ||||
Range(mm) | 5-140 | 5-140 | 15-75 | ||||
< 30 mm | 396 | 71 | 388 | 72 | 9 | 47 | < 0.001* |
≥ 30 mm | 163 | 29 | 152 | 28 | 10 | 53 | |
Pathological stage | |||||||
IA | 363 | 65 | 360 | 67 | 4 | 21 | < 0.001** |
IB | 95 | 17 | 88 | 16 | 6 | 32 | |
IIA | 36 | 6 | 31 | 6 | 5 | 26 | |
IIB | 13 | 2 | 13 | 2 | 0 | 0 | |
IIIA | 42 | 8 | 39 | 7 | 3 | 16 | |
≥ IIIB | 10 | 2 | 9 | 2 | 1 | 5 | |
Lymph node metastasis | |||||||
NX | 69 | 12 | 68 | 13 | 1 | 5 | |
N0 | 420 | 75 | 409 | 75 | 11 | 58 | 0.002** |
≥ N1 | 70 | 13 | 63 | 12 | 7 | 21 | |
Pleural invasion | |||||||
Negative | 452 | 80 | 446 | 83 | 6 | 32 | < 0.001** |
Positive | 107 | 20 | 94 | 17 | 13 | 68 | |
Lymphatic invasion | |||||||
Negative | 466 | 83 | 461 | 85 | 5 | 26 | < 0.001** |
Positive | 93 | 17 | 79 | 15 | 14 | 74 | |
Vascular invasion | |||||||
Negative | 427 | 76 | 422 | 78 | 5 | 26 | < 0.001** |
Positive | 132 | 24 | 118 | 22 | 14 | 74 |
Survival analysis
No. | % | Univariate analysis | Multivariate analysis | |||
---|---|---|---|---|---|---|
P value | Hazard ratio | 95% CI | P value | |||
Total | 559 | |||||
Age | ||||||
< 65 | 213 | 38 | 0.388 | 1.000 | ||
≥ 65 | 346 | 62 | 1.933 | 0.849-4.402 | 0.116 | |
Gender | ||||||
Female | 288 | 52 | 0.768 | 1.000 | ||
Male | 271 | 48 | 0.807 | 0.232-2.803 | 0.735 | |
Smoking status | ||||||
Non-smoker | 284 | 49 | 0.560 | 1.000 | ||
Smoker | 275 | 51 | 1.164 | 0.342-3.956 | 0.808 | |
Tumor size | ||||||
< 30 mm | 396 | 71 | 0.059 | 1.000 | ||
≥ 30 mm | 163 | 29 | 0.819 | 0.338-1.985 | 0.658 | |
Pathological stage | ||||||
I | 458 | 82 | <0.001 | 1.000 | ||
II, III, IV | 101 | 18 | 2.768 | 1.113-6.884 | 0.028 | |
Pleural invasion | ||||||
Negative | 452 | 81 | <0.001 | 1.000 | ||
Positive | 107 | 19 | 0.848 | 0.345-2.083 | 0.719 | |
Lymphatic invasion | ||||||
Negative | 466 | 83 | <0.001 | 1.000 | ||
Positive | 93 | 17 | 3.430 | 1.363-8.634 | 0.009 | |
Vascular invasion | ||||||
Negative | 427 | 76 | <0.001 | 1.000 | ||
Positive | 132 | 24 | 3.309 | 1.312-8.350 | 0.011 | |
SMPC | ||||||
Negative | 540 | 97 | <0.001 | 1.000 | ||
Positive | 19 | 3 | 1.871 | 0.528-6.630 | 0.332 | |
AMPC | ||||||
Negative | 460 | 83 | 0.045 | 1.000 | ||
Positive | 99 | 17 | 1.132 | 0.450-2.845 | 0.792 |
No. | % | Univariate Analysis | Multivariate analysis | |||
---|---|---|---|---|---|---|
P value | Hazard ratio | 95% CI | P value | |||
Total | 458 | |||||
Age | ||||||
< 65 | 172 | 38 | 0.394 | 1.000 | ||
≥ 65 | 286 | 62 | 2.191 | 0.474-10.131 | 0.316 | |
Gender | ||||||
Female | 249 | 54 | 0.063 | 1.000 | ||
Male | 209 | 46 | 0.157 | 0.014-1.787 | 0.136 | |
Smoking status | ||||||
Non-smoker | 248 | 54 | 0.204 | 1.000 | ||
Smoker | 210 | 46 | 0.768 | 0.117-5.052 | 0.784 | |
Tumor size | ||||||
< 30 mm | 358 | 78 | 0.264 | 1.000 | ||
≥ 30 mm | 100 | 22 | 0.304 | 0.037-2.504 | 0.268 | |
Pleural invasion | ||||||
Negative | 402 | 88 | < 0.001 | 1.000 | ||
Positive | 56 | 12 | 1.519 | 0.328-7.040 | 0.593 | |
Lymphatic invasion | ||||||
Negative | 415 | 91 | < 0.001 | 1.000 | ||
Positive | 43 | 9 | 5.016 | 1.295-19.434 | 0.020 | |
Vascular invasion | ||||||
Negative | 390 | 85 | < 0.001 | 1.000 | ||
Positive | 68 | 15 | 4.494 | 1.006-20.081 | 0.049 | |
SMPC | ||||||
Negative | 448 | 98 | < 0.001 | 1.000 | ||
Positive | 10 | 2 | 9.028 | 1.164-70.031 | 0.035 | |
AMPC | ||||||
Negative | 389 | 98 | 0.023 | 1.000 | ||
Positive | 69 | 2 | 1.825 | 0.378-8.808 | 0.454 |
Immunohistochemical findings
Classification/Antibody | SMPC | AMPC | nMPC |
---|---|---|---|
Cellular adhesion molecules | |||
E-cadherin | 215.3* | 143.9 | 187.1 |
CD44 | 60.8‡ | 205.9¶ | 141.3 |
Laminin5γ2 | 69.4 | 36.9 | 60.3 |
Growth factor | |||
VEGF-C | 294.4 | 296.4 | 282.1 |
Apoptosis-associated proteins | |||
bcl2 | 13.2 | 11.1 | 21.8 |
p53§ | 36.4 | 26.1 | 45.0 |
cleaved caspase-3§ | 0.3 | 0.2 | 0.4 |
Mucin-related proteins | |||
MUC1 | 169.7 | 182.5 | 202.1 |
MUC6 | 0.0 | 0.0 | 0.0 |
Hypoxia induced protein | |||
HIF-1α | 1.8 | 2.4 | 2.9 |
Others | |||
TTF-1 | 267.9 | 289.3 | 248.6 |
SP-A | 45.2‡ | 82.6 | 123.2 |
Vimentin | 112.1 | 117.9 | 72.1 |
Ki-67§ | 22.6 | 16.9 | 16.2 |
LYVE1 | 98.9 | 107.2 | 101.9 |
c-Met | 217.2 | 253.8 | 211.4 |
Phospho-c-Met | 34.2‡ | 50.0 | 88.0 |
Mutation analysis
total | % | SMPC(+) cases | % | SMPC(-) cases | % | P value | |
---|---|---|---|---|---|---|---|
No. | 33 | 19 | 14 | ||||
EGFR mutation | |||||||
Negative | 13 | 39 | 5 | 26 | 8 | 57 | 0.076* |
Positive | 20 | 61 | 14 | 74 | 6 | 43 | |
ex19 | 7 | 35§ | 5 | 36§ | 2 | 33§ | 0.664* |
ex21 | 13 | 65§ | 9 | 64§ | 4 | 67§ | |
KRAS mutation | |||||||
Negative | 33 | 100 | 19 | 100 | 14 | 100 | - |
Positive | 0 | 0 | 0 | 0 |