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Supplementary Table 1: Immunohistochemistry

Antibody Clone Vendor Titer

Chromogranin LK2H10 Ventana 1µg/mL

Synaptophysin 27G12 Leica

(Novocastra) 1:100-200

CD56 123C3 Dako 1:50-100

NSE 22C9 Leica

(Novocastra) 1:200-400

TTF1 SPT24 Leica

(Novocastra) 1:200

Ki67 MIB-1 Dako 1:75-150

p16 E6H4 Ventana 1µg/mL

p53 DO-7 Ventana 0.5µg/mL

p63 4A4 Biocare 1:100-200

c-Myc

EP121 (also known as

Y69)

Epitomics 1:100-200

CyclinD1 SP4 Thermo (Lab

Vision) 1:25-50

CD117 A4502 Dako 1:400-600

HER2 4B5 Ventana 6µg/mL

Supplementary Table 2: The results of immunostaining for markers of neuroendocrine differentiation (synaptophysin, chromogranin, NSE, CD56 and TTF1)

Neuroendocrin e

Marker

UC (N=119)

HGNE (N=76)

Small Cell (n=65)

LCNE (n=5)

LCNE+Mixed (n=11)

Chromogranin

No Staining 119 (100.0%) 18 (23.7%) 13 (20.0%) 3 (60.0%) 5 (45.5%)

1+ 0 (0.0%) 28 (36.8%) 28 (43.1%) 0 (0.0%) 0 (0.0%)

2+ 0 (0.0%) 13 (17.1%) 11 (16.9%) 0 (0.0%) 2 (18.2%)

3+ 0 (0.0%) 17 (22.4%) 13 (20.0%) 2 (40.0%) 4 (36.4%)

Positive staining 0 (0.0%) 58 (76.3%) 52 (80.0%) 2 (40.0%) 6 (54.6%)

Synaptophysin

No Staining 116 (97.5%) 6 (7.9%) 5 (7.7%) 0 (0.0%) 1 (9.1%)

1+ 3 (2.5%) 19 (25.0%) 17 (26.1%) 1 (20.0%) 2 (18.2%)

2+ 0 (0.0%) 20 (26.3%) 16 (24.6%) 2 (40.0%) 4 (36.4%)

3+ 0 (0.0%) 31 (40.8%) 27 (41.5%) 2 (40.0%) 4 (36.4%)

Positive staining 3 (2.5%) 70 (92.1%) 60 (92.2%) 5 (100.0%) 10 (91.0%)

NSE

No Staining 26 (21.8%) 0 (0.0%) 0 (0.0%) 0 (0.0%) 0 (0.0%)

1+ 72 (60.5%) 10 (13.1%) 9 (13.9%) 1 (20.0%) 1 (9.1%)

2+ 19 (16.0%) 9 (11.8%) 7 (10.8%) 1 (20.0%) 2 (18.2%)

3+ 2 (1.7%) 57 (75.0%) 49 (75.4%) 3 (60.0%) 8 (72.7%)

Positive staining 93 (78.2%) 76 (100.0%) 65 (100.0%) 5 (100.0%) 11 (100.0%)

CD56

No Staining 114 (95.8%) 6 (7.9%) 5 (7.7%) 0 (0.0%) 1 (9.1%)

1+ 2 (1.7%) 17 (22.4%) 14 (21.5%) 2 (40.0%) 3 (27.3%)

2+ 3 (2.5%) 27 (35.5%) 24 (36.9%) 1 (20.0%) 3 (27.3%)

3+ 0 (0.0%) 26 (34.2%) 22 (33.9%) 2 (40.0%) 4 (36.4%)

Positive staining 5 (4.2%) 70 (92.1%) 60 (92.3%) 5 (100.0%) 10 (91.0%)

TTF1

No Staining 115 (96.6%) 35 (46.1%) 31 (47.7%) 2 (40.0%) 4 (36.4%)

1+ 4 (3.4%) 17 (22.4%) 13 (20.0%) 2 (40.0%) 4 (36.4%)

2+ 0 (0.0%) 12 (15.8%) 10 (15.4%) 1 (20.0%) 2 (18.2%)

3+ 0 (0.0%) 12 (15.8%) 11 (16.9%) 0 (0.0%) 1 (9.1%)

Positive staining 4 (3.4%) 41 (54.0%) 34 (52.3%) 3 (60.0%) 7 (63.7%)

Statistical analysis was performed for negative and positive staining. Positive staining

was further graded as 1+ (minimal staining), 2+ (moderate staining) and 3+ (intense

staining).

Supplementary Table 3: The results of immunostaining for cell cycle regulators (p16, p53, p63, c-Myc, cyclinD1)

Cell Cycle Related Markers

UC (N=119)

HGNE (N=76)

Small Cell (n=65)

LCNE (n=5)

LCNE+Mixed (n=11)

p16

No staining 52 (43.7%) 2 (2.6%) 2 (3.1%) 0 (0.0%) 0 (0.0%)

1+ 10 (8.4%) 1 (1.3%) 1 (1.5%) 0 (0.0%) 0 (0.0%)

2+ 14 (11.8%) 5 (6.6%) 4 (6.1%) 1 (20.0%) 1 (9.1%)

3+ 43 (36.1%) 68 (89.5%) 58 (89.2%) 4 (80.0%) 10 (91.0%)

Positive staining 67 (56.3%) 74 (97.4%) 63 (96.8%) 5 (100.0%) 11 (100.0%) p53

No staining 17 (14.3%) 7 (9.2%) 6 (9.2%) 1 (20.0%) 1 (9.1%)

1+ 19 (16.0%) 4 (5.3%) 4 (6.2%) 0 (0.0%) 0 (0.0%)

2+ 27 (22.7%) 15 (19.7%) 12 (18.5%) 3 (60.0%) 3 (27.3%)

3+ 56 (47.1%) 50 (65.8%) 43 (66.2%) 1 (20.0%) 7 (63.7%)

Positive staining 102 (85.7%) 69 (90.8%) 59 (90.9%) 4 (80.0%) 10 (91.0%) p63

No staining 10 (8.4%) 38 (50.0%) 33 (50.8%) 3 (60.0%) 5 (45.5%)

1+ 12 (10.1%) 27 (35.5%) 24 (36.9%) 0 (0.0%) 3 (27.3%)

2+ 25 (21.0%) 5 (6.6%) 4 (6.2%) 1 (20.0%) 1 (9.1%)

3+ 72 (60.5%) 6 (7.9%) 4 (6.2%) 1 (20.0%) 2 (18.2%)

Positive staining 109 (91.6%) 38 (50.0%) 32 (49.3%) 2 (40.0%) 6 (54.6%) c-Myc

No staining 28 (23.5%) 38 (50.0%) 33 (50.8%) 3 (60.0%) 5 (45.5%)

1+ 61 (51.3%) 31 (40.8%) 28 (43.1%) 0 (0.0%) 3 (27.3%)

2+ 29 (24.4%) 6 (7.9%) 3 (4.6%) 2 (40.0%) 3 (27.3%)

3+ 1 (0.8%) 1 (1.3%) 1 (1.5%) 0 (0.0%) 0 (0.0%)

Positive staining 91 (76.5%) 38 (50.0%) 32 (49.2%) 2 (40.0%) 6 (54.6%) CyclinD1

No staining 6 (5.0%) 37 (48.7%) 32 (49.2%) 2 (40.0%) 5 (45.5%)

1+ 22 (18.5%) 27 (35.5%) 23 (35.4%) 1 (20.0%) 4 (36.4%)

2+ 43 (36.1%) 10 (13.2%) 8 (12.3%) 2 (40.0%) 2 (18.2%)

3+ 48 (40.3%) 2 (2.7%) 2 (3.1%) 0 (0.0%) 0 (0.0%)

Positive staining 113 (95.0%) 39 (51.4%) 33 (50.8%) 3 (60.0%) 6 (54.6%)

Statistical analysis was performed for negative and positive staining. Positive staining

was further graded as 1+ (minimal staining), 2+ (moderate staining) and 3+ (intense

staining).

Supplementary Table 4: The results of immunostaining for markers of targeted therapies (CD117 and HER2)

Targeted Therapy Related

UC (N=119)

HGNE (N=76)

Small Cell (n=65)

LCNE (n=5)

LCNE+Mixed (n=11)

HER2

No staining 34 (28.6%) 63 (82.9%) 55 (84.6%) 3 (60.0%) 8 (72.8%)

1+ 39 (32.8%) 5 (6.6%) 4 (6.2%) 1 (20.0%) 1 (9.1%)

2+ 32 (26.9%) 4 (5.3%) 2 (3.1%) 1 (20.0%) 2 (18.2%)

3+ 14 (11.8%) 4 (5.3%) 4 (6.2%) 0 (0.0%) 0 (0.0%)

Positive staining 85 (71.4%) 13 (17.2%) 10 (15.4%) 2 (40.0%) 3 (27.3%) CD117

No staining 101 (84.9%) 28 (36.9%) 24 (36.9%) 2 (40.0%) 4 (36.4%)

1+ 16 (13.4%) 29 (38.1%) 27 (41.5%) 1 (20.0%) 2 (18.2%)

2+ 2 (1.7%) 16 (21.1%) 11 (16.9%) 2 (40.0%) 5 (45.5%)

3+ 0 (0.0%) 3 (3.9%) 3 (4.6%) 0 (0.0%) 0 (0.0%)

Positive staining 18 (15.1%) 48 (63.1%) 41 (63.1%) 3 (60.0%) 7 (63.7%)

Statistical analysis was performed for negative and positive staining. Positive staining

was further graded as 1+ (minimal staining), 2+ (moderate staining) and 3+ (intense

staining).