CD24 genetic variants contribute to overall survival in patients with gastric cancer

Table 1 Primer sequences used for polymerase chain reaction

Primer	Sequence	Length of product	Endonucleases	Bands
P-534F	5’-AGAGATAACCCTGCCCGAG-3’	209 bp	BsrFI	C: 126 bp + 83 bp
P-534R	5’-CCAAGTTTCCTTTGTTTCCC-3’
outerF	5’-CCACTTGGCATTTTTGAGGCATCT-3’	1882 bp	-	-
outerR	5’-TGTGTCGAGGCAGTTGTAAAAG-3’
P170F	5’-CTAAAGAGAATGACCTTGGTGGGTTGAG-3’	404 bp	BstXI	T: 275 bp + 129 bp
P170R	5’-GGATTGGGTTTAGAAGATGGGGAAA-3’
P1527F	5’-GCCAGGGCAATGATGAATGAG-3’	847 bp	BsrI	TG: 645 bp + 202 bp
P1527R	5’-TGTGTCGAGGCAGTTGTAAAAGAT-3’

Table 2 Characteristics of subjects included n (%)

	Cancer	Atrophy	Controls	P value
n	679	111	976
Gender
Male	487 (71.7)	66 (59.5)	578 (59.2)	< 0.0001
Female	192 (28.3)	45 (40.5)	398 (40.8)
Age (yr)	61.0 (54.0-70.0)	50.0 (47.0-57.0)	48.5 (44.0-55.0)	< 0.0001
≤ 55	204 (30.0)	76 (68.5)	756 (77.5)	< 0.0001
55-70	310 (45.7)	28 (25.2)	181 (18.5)
> 70	165 (24.3)	7 (6.3)	39 (4.0)
H. pylori
Positive	453 (67.8)	84 (75.7)	478 (49.7)	< 0.0001
Negative	215 (32.2)	27 (24.3)	483 (50.3)
Differentiation
Poor	437 (68.4)
Moderate to well	202 (31.6)
Pathologic type
Tubular adenocarcinoma	545 (82.2)
Signet ring cell	50 (7.5)
Other	68 (10.3)
TNM stage
I	97 (15.0)
II	227 (35.1)
III	224 (34.6)
IV	99 (15.3)
Distant metastasis
Positive	573 (85.3)
Negative	99 (14.7)
Post-operational Chemotherapy
No	456 (67.2)
FOLFOX-4	100 (14.7)
XELOX	47 (6.9)
Other	76 (11.2)

Data are presented as frequency counts (percentage of total) or median (25^th to 75^th percentiles).

Table 3 Distributions of genotypes in three groups n (%)

	Controls	Cancer	P value	OR1 (95%CI)	Atrophy	P value	OR1 (95%CI)
P-534
C/C	271 (27.8)	181 (26.7)	-	Reference	29 (26.1)	-	Reference
C/A	488 (50.0)	358 (52.7)	0.099	1.1 (0.86-1.48)	62 (55.9)	0.204	1.2 (0.74-1.91)
A/A	217 (22.2)	140 (20.6)	0.145	0.9 (0.61-1.20)	20 (18.0)	0.328	0.8 (0.46-1.54)
C	1030 (52.8)	720 (53.0)	0.886	-	130 (54.1)	0.716	-
A	922 (47.2)	638 (47.0)		1.0 (0.86-1.14)	102 (45.9)		0.9 (0.72-1.25)
P170
C/C	419 (42.9)	295 (43.4)	-	Reference	60 (54.1)	-	Reference
C/T	439 (45.0)	308 (45.4)	0.277	1.1 (0.82-1.38)	37 (33.3)	0.082	0.6 (0.40-0.96)
T/T	118 (12.1)	76 (11.2)	0.369	0.9 (0.60-1.29)	14 (12.6)	0.724	0.9 (0.47-1.65)
C	1277 (65.4)	898 (66.1)	0.673	-	157 (70.7)	0.114	-
T	675 (34.6)	460 (33.9)		1.0 (0.83-1.12)	65 (29.3)		0.8 (0.58-1.06)
P1527
TG/TG	826 (84.6)	559 (82.3)	-	Reference	95 (85.6)	-	Reference
TG/del	142 (14.6)	117 (17.2)	0.069	1.3 (0.97-1.82)	15 (13.5)	0.940	0.9 (0.50-1.59)
del/del	8 (0.8)	3 (0.4)	0.162	0.4 (0.09-1.77)	1 (0.9)	0.935	0.9 (0.10-7.30)
TG	1794 (91.9)	1235 (90.9)	0.956	-	205 (92.3)	0.821	-
del	158 (8.1)	123 (9.1)		1.1 (0.88-1.45)	17 (7.7)		0.9 (0.56-1.58)
Haplotype2
ACTG	45.6	47.0	-	Reference	45.4	-	Reference
CTTG	33.6	33.7	0.946	1.0 (0.85-1.16)	28.7	0.407	0.9 (0.63-1.21)
CCTG	11.4	10.2	0.382	0.9 (0.71-1.14)	17.7	0.072	1.6 (0.99-2.37)
CCdel	8.0	9.1	0.370	1.1 (0.87-1.46)	7.7	0.919	1.0 (0.56-1.67)

Data are presented as frequency counts (percentage of total).

¹ORs of the genotypes were calculated adjusting for age, sex and H. pylori infection in logistic regression model;

²The haplotype was lined with P-534, P170 and P1527 and displayed as percentage.

Table 4 Distributions of genotypes according to clinical parameters in gastric cancer cases

	P-534			P value	P170			P value	P1527		P value
	C/C	C/A	A/A		C/C	C/T	T/T		TG/TG	TG/del
n	181	358	140		295	308	76		559	120
Age	60 (53-70)	61 (54-71)	61 (55-70)	0.715	61 (53-71)	61 (54-70)	63 (56-70)	0.658	61 (54-70)	60 (51-71)	0.307
Sex
Male	27.3	50.3	22.4	0.092	43.9	44.2	11.9	0.485	83.6	16.4	0.172
Female	25.0	58.9	16.1		42.2	48.4	9.4		79.2	20.8
H. pylori
Positive	26.3	53.4	20.3	0.905	42.6	46.1	11.3	0.769	82.3	17.7	0.996
Negative	27.0	51.6	21.4		45.6	43.7	10.7		82.3	17.7
Differentiation
Poor	24.7	55.8	19.5	0.070	43.5	47.4	9.2	0.055	81.5	18.5	0.500
Moderate to well	30.2	46.0	23.8		45.0	40.1	14.8		83.7	16.3
TNM stage
I-II	26.2	52.8	21.0	0.999	45.1	44.7	10.2	0.695	82.4	17.6	0.986
III-IV	26.3	52.6	21.1		42.7	45.2	12.1		82.4	17.6
Pathologic type
Tubular adenocarcinoma	26.4	52.3	21.3	0.754	45.0	43.9	11.2	0.173	82.8	17.2	0.751
Signet ring cell	26.0	60.0	14.0		34.0	60.0	6.0		80.0	20.0
Other	27.9	50.0	22.1		38.2	47.1	14.7		85.3	14.7
Distant metastasis
Positive	32.3	48.5	19.2	0.323	40.4	44.4	15.1	0.374	79.8	20.2	0.454
Negative	25.1	53.9	20.9		44.3	45.2	10.5		82.9	17.1
CD24 staining	60 (0-120)	60 (0-120)	100 (20-140)	0.215	60 (0-120)	60 (20-120)	40 (0-100)	0.482	60 (0-120)	60 (40-100)	0.922

Data are presented as percentage of total or median (25^th to 75^th percentiles).

Table 5 Results of multivariate Cox regression analysis

	P value	HR	95%CI
P-534
C/C + C/A	-	1.00	-
A/A	0.0416	1.38	1.01-1.88
Age	0.1875	1.01	1.00-1.02
Sex
Male	-	1.00	-
Female	0.2315	0.83	0.61-1.13
Differentiation
Moderate to well	-	1.00	-
Poor	0.0089	1.50	1.11-2.04
Pathologic type
Tubular adenocarcinoma	-	1.00	-
Signet ring cell	0.4750	0.83	0.50-1.39
Other	0.9469	1.01	0.67-1.53
TNM stage
I	-	1.00	-
II	0.0003	5.50	2.18-13.86
III	< 0.0001	22.29	9.05-54.89
IV	< 0.0001	32.12	12.63-81.70
Chemotherapy
No	-	1.00	-
FOLFOX-4	0.0195	0.64	0.45-0.93
XELOX	0.0021	0.43	0.25-0.74
Other	0.0004	0.45	0.29-0.70