Phenotypic classification of gastric signet ring cell carcinoma and its relationship with clinicopathologic parameters and prognosis

Table 1 Antibodies for immunohistochemistry

Antibody	Clone	Diluent’sproportion	Company
Gastric phenotypic markers
MUC5AC	CLH2	1:100	Novocastra, Newcastle, UK
MUC6	CLH5	1:50	Novocastra, Newcastle, UK
HGM	45M1	1:50	Novocastra, Newcastle, UK
Intestinal phenotypic markers
MUC2	Ccp58	1:100	Novocastra, Newcastle, UK
CDX2	CDX2-88	1:50	BioGenex, San Ramon CA
Li-cadherin	Clone 141713	1:200	Neomarker, Fremont, CA, USA
VILLIN	CWWB1	1:200	Novocastra, Newcastle, UK
Other antibodies
Ki -67	K-2	1:50	Zymed, San Francisco, CA, USA
CD44v6	VFF-7	1:50	Zymed , San Francisco, CA, USA
E-cadherin	4A2C7	1:50	Zymed , San Francisco, CA, USA
β-catenin	CAT-5H10	1:30	Zymed , San Francisco, CA, USA

Table 2 Gastric and intestinal phenotypic marker expression in gastric SRC carcinoma

Phenotype	Age/Gender	MUC2	Li-cadherin	CDX2	VILLIN	HGM	MUC5AC	MUC6
G type (n = 17)
1	65/F	-	-	-	-	+	+	+
2	60/M	-	-	-	-	+	+	-
3	67/F	-	-	-	-	+	+	-
4	29/F	-	-	-	-	+	+	+
5	66/F	-	-	-	-	+	+	+
6	49/M	-	-	-	-	-	+	-
7	49/M	-	-	-	-	+	+	-
8	69/F	-	-	-	-	+	+	-
9	33/F	-	-	-	-	+	+	-
10	38/F	-	-	-	-	-	+	-
11	35/F	-	-	-	-	+	+	-
12	56/M	-	-	-	-	+	+	-
13	39/M	-	-	-	-	-	+	-
14	70/F	-	-	-	-	+	+	-
15	42/M	-	-	-	-	+	+	-
16	42.M	-	-	-	-	-	+	-
17	66/M	-	-	-	-	+	-	-
I type (n = 10)
1	63/M	+	+	+	-	-	-	-
2	75/M	+	+	+	-	-	-	-
3	59/F	+	+	+	-	-	-	-
4	60/M	+	-	-	-	-	-	-
5	31/F	+	-	-	-	-	-	-
6	76/F	+	-	-	-	-	-	-
7	73/F	+	-	-	+	-	-	-
8	46/F	+	+	-	-	-	-	-
9	62/M	-	+	-	+	-	-	-
10	41/M	-	+	-	-	-	-	-
GI type (n = 31)
1	31/M	+	-	-	-	+	+	-
2	69/F	+	-	-	-	+	+	-
3	61/M	+	-	+	-	+	+	+
4	67/M	+	-	-	-	+	+	+
5	56/M	+	+	+	-	+	+	-
6	47/F	+	+	-	-	+	+	-
7	65/M	+	+	-	-	+	+	-
8	64/M	+	-	-	-	+	+	-
9	63/M	+	+	+	-	+	+	-
10	48/F	+	+	-	-	+	+	+
11	52/F	+	-	-	-	+	+	-
12	65/M	+	+	+	-	+	+	-
13	72/M	+	+	-	-	+	+	-
14	39/F	+	+	+	-	+	+	-
15	36/M	+	+	+	+	+	+	-
16	51/M	+	+	-	-	+	+	-
17	60/F	+	+	-	-	+	-	-
18	46/F	+	-	-	-	+	-	-
19	45/F	+	-	-	-	+	-	-
20	45/F	+	-	+	-	+	-	-
21	55/M	+	+	+	-	+	-	-
22	65/F	+	-	-	-	+	-	-
23	47/F	+	-	-	-	+	-	-
24	60/F	+	+	-	-	+	-	-
25	46/F	-	-	+	-	+	+	+
26	50/F	-	+	-	-	+	+	-
27	54/F	-	+	+	-	+	+	-
28	32/M	-	-	-	+	+	+	-
29	55/M	-	+	+	+	+	+	-
30	38/F	-	+	-	-	+	-	-
31	59/M	-	+	-	-	+	-	-
U type (n = 8)
1	36/M	-	-	-	-	-	-	-
2	49/F	-	-	-	-	-	-	-
3	39/M	-	-	-	-	-	-	-
4	61/F	-	-	-	-	-	-	-
5	70/F	-	-	-	-	-	-	-
6	67/M	-	-	-	-	-	-	-
7	64/M	-	-	-	-	-	-	-
8	34/F	-	-	-	-	-	-	-

Table 3 Different expression patterns of phenotype markers and Ki67 and β-catenin expression in gastric SRC carcinoma

					β-catenin	Plasma/Nucleusβ-cateninexpression
Phenotype of gastricSRC carcinomas	Number ofpositive GPM	Number ofpositive IPM	Cases	Ki67 index> 25%	membraneexpression
G type			17	2 (11.8%)	11 (64.7%)	2 (11.8%)
G1	1	0	5	0	3 (60.0%)	0
G2	2	0	9	2 (22.2%)	6 (66.7%)	1 (11.1%)
G3	3	0	3	0	2 (66.7%)	1 (33.3%)
I type			10	5 (50.0%)	1 (10.0%)b	4 (40.0%)
I1	0	1	4	1 (25.0%)	0	1 (25.0%)
I2	0	2	3	3 (100%)	0	2 (66.7%)
I3	0	3	3	1 (33.3%)	1 (33.3%)	1 (33.3%)
GI type			31	15 (48.4%)a	12 (36.4%)	14 (45.2%)a
GI1	1	1	6	3 (50.0%)	2 (33.3%)	3 (50.0%)
GI2	1	2	3	1 (33.3%)	1 (33.3%)	1 (33.3%)
GI3	1	3	1	0	0	1 (100%)
GI4	2	1	6	3 (50.0%)	3 (50.0%)	3 (50.0%)
GI5	2	2	5	4 (80.0%)	4 (80.0%)	2 (40.0%)
GI6	2	3	5	2 (40.0%)	1 (20.0%)	1 (20.0%)
GI7	2	4	1	0	0	1 (100%)
GI8	3	1	2	1 (50.0%)	1 (50.0%)	1 (50.0%)
GI9	3	2	2	1 (50.0%)	0	1 (50.0%)
U type	0	0	8	3 (37.5%)	4 (50.0%)	3 (37.5%)

GPM: Gastric phenotype marker. IPM: Intestinal phenotype marker.

^aP < 0.05,

^bP < 0.05 vs G type.

Table 4 Correlation between muc2, cdx2, Li-cadherin, Ki-67 index and clinicopathologic features n (%)

		MUC2 expression			CDX2 expression			Li-cadherin expression			Ki-67 index
Factors	Cases	MUC2-(n = 34)	MUC2+(n = 32)	P	CDX2-(n = 52)	CDX2+(n = 14)	P	Li-cad-(n = 42)	Li-cad+(n = 24)	P	≤ 25%(n = 40)	> 25%(n = 26)	P
Gender
Male	32	17 (50.0)	15 (46.9)	> 0.05	23 (44.2)	9 (64.3)	> 0.05	18 (42.9)	14 (58.3)	> 0.05	18 (45.0)	14 (53.8)	> 0.05
Female	34	17 (50.0)	17 (53.1)		29 (55.8)	5 (35.7)		24 (57.1)	10 (41.7)		22 (55.0)	12 (46.2)
Age (yr)
Mean ± SE	66	50.9 ± 2.2	56.0 ± 2.2	> 0.051	53.5 ± 1.9	52.9 ± 3.1	> 0.051	51.9 ± 3.0	54.6 ± 2.6	> 0.051	53.9 ± 2.2	51.0 ± 2.3	> 0.051
Tumor diameter
< 5.0 cm	32	23 (67.6)	9 (28.1)	0.001	30 (57.7)	2 (14.3)	0.004	24 (57.1)	8 (33.3)	> 0.05	24 (60.0)	8 (30.8)	0.02
≥ 5.0 cm	34	11 (32.4)	23 (71.9)		22 (42.3)	12 (85.7)		18 (42.9)	16 (66.7)		16 (40.0)	18 (69.2)
Depth of wall invasion
T1	13	11 (32.4)	2 (6.3)	0.0082	13 (25.0)	0	> 0.05	10 (23.8)	3 (12.5)	> 0.05	11 (27.5)	2 (7.7)	0.0482
T2	22	10 (29.4)	12 (37.5)		16 (30.8)	6 (42.9)		15 (35.7)	7 (29.2)		14 (35.0)	8 (30.8)
T3	22	10 (29.4)	12 (37.5)		15 (28.8)	7 (50.0)		11 (26.2)	11 (45.8)		10 (25.0)	12 (46.2)
T4	9	3 (8.8)	6 (18.8)		8 (15.4)	1 (7.1)		6 (14.3)	3 (12.5)		5 (12.5)	4 (15.4)
Lymph node metastasis
LN (–)	29	21 (61.8)	8 (25.0)	0.003	25 (48.1)	4 (28.6)	> 0.05	20 (47.6)	9 (37.5)	> 0.05	22 (55.0)	7 (26.9)	0.025
LN (+)	37	13 (38.2)	24 (75.0)		27 (51.9)	10 (71.4)		22 (52.4)	15 (62.5)		18 (45.0)	19 (70.1)
Distant metastasis
M0	60	32 (94.1)	28 (87.5)	> 0.05	48 (92.3)	12 (85.7)	> 0.05	39 (92.9)	21 (87.5)	> 0.05	36 (90.0)	24 (92.3)	> 0.05
M1	6	2 (5.9)	4 (12.5)		4 (7.7)	2 (14.3)		3 (7.1)	3 (12.5)		4 (10.0)	2 (7.7)
Vascular invasion
V (–)	40	27 (79.4)	13 (40.6)	0.001	34 (65.4)	6 (42.9)	> 0.05	31 (73.8)	9 (37.5)	0.004	28 (70.0)	12 (46.2)	> 0.05
V (+)	26	7 (20.6)	19 (59.4)		18 (34.6)	8 (57.1)		11 (26.2)	15 (62.5)		12 (30.0)	14 (53.8)
TNM stage
I+ II	32	22 (64.7)	10 (31.2)	0.007	27 (51.9)	5 (35.7)	> 0.05	21 (50.0)	11 (45.8)	> 0.05	24 (60.0)	8 (30.8)	0.02
III + IV	34	12 (35.3)	22 (68.8)		25 (48.1)	9 (64.3)		21 (50.0)	13 (54.2)		16 (40.0)	18 (69.2)

¹t test;

²T1 vs T2 + T3 + T4.

Table 5 Correlation between four phenotypes of gastric SRC carcinoma and clinicopathologic features n (%)

		Four phenotypes of gastric SRC carcinoma
Factors	n	G type (n = 17)	I type (n = 10)	GI type (n = 31)	UC type (n = 8)
Gender
Male	32	8 (47.1)	5 (50.0)	15 (48.4)	4 (50.0)
Female	34	9 (52.9)	5 (50.0)	16 (51.6)	4 (50.0)
Age (yr)
Mean ± SE	66	53.7 ± 2.9	49.1 ± 4.0	54.7 ± 2.5	52.8 ± 4.3
Tumor diameter
< 5.0 cm	32	15 (88.2)a	3 (30.0)b	10 (32.3)	4 (50.0)
≥ 5.0 cm	34	2 (11.8)a	7 (70.0)b	21 (67.7)	4 (50.0)
Depth of wall invasion
T1	13	7 (41.2)c	3 (30.0)	1 (3.2)	2 (25.0)
T2	22	5 (29.1)c	2 (20.0)	12 (38.7)	3 (37.5)
T3	22	4 (23.5)c	2 (20.0)	13 (41.9)	3 (37.5)
T4	9	1 (5.9)c	3 (30.0)	5 (16.1)	0
Lymph node metastasis
LN (–)	29	13 (76.5)c	4 (40.0)	8 (25.8)	4 (50.0)
LN (+)	37	4 (23.5)c	6 (60.0)	23 (74.2)	4 (50.0)
Distant metastasis
M0	60	16 (94.1)	8 (80.0)	28 (90.3)	8 (100)
M1	6	1 (5.9)	2 (20.0)	3 (9.7)	0
Vascular invasion
V (–)	40	15 (88.2)c	7 (70.0)	12 (38.7)	6 (75.0)
V (+)	26	2 (11.8)c	3 (30.0)	19 (61.3)	2 (25.0)
TNM stage
I+ II	32	12 (70.6)d	4 (40.0)	12 (38.7)	4 (50.0)
III + IV	34	5 (29.1)d	6 (60.0)	19 (61.3)	4 (50.0)
IM
IM (–)	46	11 (64.7)	6 (60.0)	23 (74.2)	6 (75.0)
IM (+)	20	6 (35.3)	4 (40.0)	8 (25.8)	2 (25.0)

IM: Intestinal metaplasia in the surrounding mucosa.

^aP < 0.001 vs GI type;

^bP < 0.01 vs I type;

^cP < 0.01 vs GI type;

^dP < 0.05 vs GI type.