Threonine and tyrosine kinase may serve as a prognostic biomarker for gallbladder cancer

Table 1 Clinicopathological characteristics of the cohort

Characteristic	n (%)
Age (yr)
≤ 65	33 (48.5)
> 65	35 (51.5)
Gender
Female	39 (57.4)
Male	29 (42.6)
Cholecystolithiasis
Yes	41 (60.3)
No	27 (39.7)
Diabetes
Yes	17 (25.0)
No	51 (75.0)
Fever
Yes	9 (13.2)
No	59 (86.8)
Jaundice
Yes	15 (22.1)
No	53 (77.9)
ALT
Normal	53 (77.9)
Elevated	15 (22.1)
AST
Normal	50 (75.8)
Elevated	16 (24.2)
TBil
Normal	49 (72.1)
Elevated	19 (27.9)
DBil
Normal	50 (73.5)
Elevated	18 (26.5)
GGT
Normal	43 (69.4)
Elevated	19 (30.6)
ALP
Normal	45 (72.6)
Elevated	17 (27.4)
CEA
Normal	42 (72.4)
Elevated	16 (27.6)
CA 19-9
Normal	26 (44.1)
Elevated	33 (55.9)
Surgical type
Curative	35 (51.5)
Noncurative	33 (48.5)
Tumor size (cm)
≤ 3	43 (63.2)
> 3	25 (36.8)
Differentiation
Lowly-undifferentiated	15 (22.1)
Moderately-well	53 (77.9)
T stage
Tis	1 (1.5)
T1	3 (4.4)
T2	29 (42.6)
T3	35 (51.5)
N stage
N0	38 (55.9)
N1	22 (32.4)
N2	8 (11.7)
M stage
M0	60 (88.2%)
M1	8 (11.8%)
TNM stage
I	4 (5.9)
II	24 (35.3)
IIIA	9 (13.2)
IIIB	17 (25.0)
IVA	0 (0)
IVB	14 (20.6)

ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; TBil: Total bilirubin; DBil: Direct bilirubin; GGT: Gamma-glutamyl transpeptidase; ALP: Alkaline phosphatase; CEA: Carcinoembryonic antigen; CA 19-9: Carbohydrate antigen 19-9.

Table 2 Threonine and tyrosine kinase staining results in tumor and normal tissues

Localization	Positive cell rate (%)		n (%)
	Median (range)
	Tumor tissues	Normal tissues	Tumor tissues	Normal tissues
Cytoplasmic staining
Negative	-	-	0	0
Positive	100 (10-100)	99 (40-100)	68 (100)	68 (100)
1+	55 (10-100)	85 (18-100)	41 (60.3)	14 (20.6)
2+	90 (5-100)	100 (80-100)	49 (72.1)	54 (79.4)
3+	20	90	1 (1.5)	1 (1.5)
Nuclear staining
Negative	-	-	8 (11.8)	2 (2.9)
Positive	15 (1-98)	60 (1-90)	60 (88.2)	66 (97.1)
1+	15 (1-85)	60 (1-90)	57 (83.8)	59 (86.8)
2+	45 (5-98)	90 (60-90)	8 (11.8)	7 (10.3)
3+	-	-	0	0

Table 3 H-score values in tumor and normal tissues

Group	n	Localization	Minimum	Maximum	Median
Tumor	68	Nucleus	0	196	12.5
	68	Cytoplasm	10	220	170
Normal	68	Nucleus	0	180	60
	68	Cytoplasm	40	270	190

Table 4 Various comparisons in tumor and normal tissues for threonine and tyrosine kinase expression

Group	n	Minimum	Maximum	Median	Mean rank	P value
Comparison between nucleus and cytoplasm in normal tissues
Nucleus	68	0	180	60	39.65	<0.0011
Cytoplasm	68	40	270	190	97.35
Comparison between nucleus and cytoplasm in tumor tissues
Nucleus	68	0	196	12.5	38.98	<0.0011
Cytoplasm	68	10	220	170	98.02
Comparison between tumor and normal tissues in nucleus
Tumor	68	0	196	12.5	49.54	<0.0011
Normal	68	0	180	60	87.46
Comparison between tumor and normal tissues in cytoplasm
Tumor	68	10	220	170	61.19	0.0261
Normal	68	40	270	190	75.81

¹Statistically significant.

Table 5 Univariate and multivariate analyses for gallbladder cancer

	χ2	P value	OR (95%CI)
Univariate
Age	2.221	0.136
Gender	0.167	0.683
Cholecystolithiasis	0.346	0.558
Diabetes	0.165	0.685
Fever	0.001	0.989
Jaundice1	5.110	0.0241
ALT1	7.781	0.0051
AST1	5.708	0.0171
TBil	0.241	0.516
DBil1	6.645	0.0101
GGT	0.899	0.343
ALP1	4.099	0.0431
CEA	3.137	0.077
CA 19-91	12.385	< 0.0011
Surgical type1	20.715	< 0.0011
Tumor size	0.099	0.754
Differentiation1	12.385	< 0.0011
T stage1	21.594	< 0.0011
N stage1	19.887	< 0.0011
M stage1	29.503	< 0.0011
TNM stage1	33.062	< 0.0011
TTK1	21.226	< 0.0011
Multivariate
Surgery type1		0.0011	4.250 (1.867-9.674)
T stage1		0.0131	2.927 (1.258-6.808)
TTK1		0.0011	0.076 (0.024-0.241)

¹Statistically significant. ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; GGT: Gamma-glutamyl transpeptidase; CEA: Carcinoembryonic antigen; TTK: Threonine and tyrosine kinase.

Table 6 Correlations between TTK expression and clinicopathological parameters

Parameter	TTK expression		χ2	P-value
	Low expression	High expression
Age (yr)
≤ 65	16	17	0.059	0.808
> 65	18	17
Gender
Female	20	19	0.277	0.598
Male	13	16
CEA
Normal	20	22	3.512	0.061
Elevated	12	4
CA 19-91
Normal	10	16	5.756	0.0161
Elevated	23	10
Tumor size
≤ 3	22	21	0.063	0.801
> 3	12	13
Differentiation1
Lowly-undifferentiated	11	4	4.191	0.0411
Moderately-well	23	30
T stage1
1 + 2	8	25	17.015	< 0.0011
3	26	9
Nodal involvement1
Negative	12	26	11.691	< 0.0011
Positive	22	8
Metastasis1
M0	27	33	5.100	0.0241
M1	7	1
TNM stage 1
I + II	5	23	19.671	< 0.0011
III + IV	29	11

¹Statistically significant. CEA: Carcinoembryonic antigen; TTK: Threonine and tyrosine kinase.