2.1 Expression of CA-IX in bladder-related tissues
Immunohistochemistry showed that CA-IX was expressed on the cell membrane, but the intracellular expression was not obvious. The positive rate of CA-IX in bladder urothelial carcinoma was 68.1% (132/194), the positive rate in adjacent tissues was 44.3% (186/194), and the positive rate in normal tissues was 27.3% (53/194) . The expression of CA-IX in bladder urothelial carcinoma was significantly higher than that in adjacent tissues and normal tissues (P<0.05). The expression intensity of CA-IX in bladder urothelial carcinoma tissue was significantly higher than that in adjacent tissue and normal tissue. See Table 1 and Figure A-C for details.
Table 1: Expression of CA-IX in bladder urothelial carcinoma tissues and other tissues
group
|
CA-Ⅸ
|
c2value
|
Pvalue
|
Positive
|
Negative
|
cancer tissue
|
132
|
62
|
|
|
Paracancer tissue
|
86
|
108
|
22.15
|
0.00
|
normal tissue
|
53
|
141
|
64.48
|
0.00
|
2.2 General clinical information
During the study period, a total of 245 patients were enrolled, and all patients had their tumors resected successfully, and the specimens were confirmed by the pathology department. A total of 194 cases were followed up for 5 years, 42 cases were lost to follow-up, and 9 cases were incomplete information. There were 145 males and 49 females; 76 cases were <60 years old, 118 cases were ≥60 years old, with an average age of 62.3±12.2 years old; 105 cases were <T1, 89 were T1; There are 80 cases of high differentiation and 114 cases of poor differentiation; 83 cases of single tumor and 111 cases of multiple tumors; 72 cases of tumor diameter ≥3cm, 122 cases of tumor diameter <3cm; 76 cases had at least one recurrence during the 5-year follow-up period, and 118 cases had no recurrence; CA-IX expression was positive in 132 cases and negative in 62 cases. According to the expression of CA-IX, the research subjects were divided into positive group and negative group, and the age, gender, T stage, degree of differentiation, tumor number, tumor diameter, and recurrence status of the two observation groups were counted.
The expression of CA-IX had no statistical significance with age, gender and tumor diameter (P>0.05); the expression of CA-IX had statistical significance with T stage, degree of differentiation, tumor number and recurrence (P<0.05). See Table 2 for details.
Table 2: Clinically relevant parameters
parameter
|
Number of cases
|
CA-Ⅸ
|
c2 value
|
P value
|
Positive(132)
|
Negative(62)
|
|
|
total
|
194
|
|
|
|
age
|
|
|
0.842
|
0.359
|
<60 years old
|
76
|
55
|
21
|
|
|
≥60 years old
|
118
|
77
|
41
|
|
|
gender
|
|
|
0.015
|
0.904
|
male
|
145
|
99
|
46
|
|
|
Female
|
49
|
33
|
16
|
|
|
T stage
|
|
|
10.412
|
0.001
|
<T1
|
105
|
61
|
44
|
|
|
T1
|
89
|
71
|
18
|
|
|
Differentiation
|
|
|
5.405
|
0.02
|
Highly
|
80
|
47
|
33
|
|
|
poorly
|
114
|
85
|
29
|
|
|
number of tumors
|
|
|
5.485
|
0.019
|
Single
|
83
|
64
|
19
|
|
|
multiple
|
111
|
68
|
43
|
|
|
tumor diameter
|
|
|
0.920
|
0.337
|
≥3cm
|
72
|
52
|
20
|
|
|
<3cm
|
122
|
80
|
42
|
|
|
recurrence
|
|
|
5.258
|
0.022
|
Yes
|
76
|
59
|
17
|
|
|
No
|
118
|
73
|
45
|
|
|
2.3 Univariate Logistic regression analysis of bladder urothelial carcinoma recurrence.
The study cases were divided into the recurrence group and the non-recurrence group according to the recurrence status, and the relevant clinical characteristics of the two study groups were summarized and analyzed. By incorporating the relevant parameters into the univariate Logistic regression analysis, we can draw the following conclusions.
Bladder urothelial carcinoma recurrence after TURBT had statistical significance (P<0.05) with T stage (OR=0.41, 95%Cl: 0.227-0.739, P=0.003), degree of differentiation (OR=0.511, 95%Cl: 0.279-0.935, P=0.029) , CA-IX expression (OR=2.396, 95%Cl: 1.233~4.654, P=0.01). Bladder urothelial carcinoma recurrence after TURBT had no statistical significance (P>0.05)with age (OR=0.895, 95%Cl: 0.496-1.613, P=0.713), gender (OR=1.227, 95%Cl: 0.636-2.369, P=0.542), tumor number (OR=0.556, 95%Cl: 0.307-1.010, P=0.054) and tumor diameter (OR=1.555, 95%Cl: 0.858-2.816, P=0.145) (P>0.05). (See Table 3 for details)
Table 3: Univariate logistic regression analysis of bladder urothelial carcinoma recurrence:
parameter
|
OR
|
95% confidence interval for EXP(B)
|
P value
|
lower limit
|
upper limit
|
age
|
0.895
|
0.496
|
1.613
|
0.712
|
gender
|
1.227
|
0.636
|
2.369
|
0.542
|
T stage
|
0.41
|
0.227
|
0.739
|
0.003
|
Differentiation
|
0.511
|
0.279
|
0.935
|
0.029
|
number of tumors
|
0.556
|
0.307
|
1.010
|
0.054
|
tumor diameter
|
1.555
|
0.858
|
2.816
|
0.145
|
CA-IX expression
|
2.396
|
1.233
|
4.654
|
0.01
|
2.4 Multivariate Logisti regression analysis of bladder urothelial carcinoma recurrence
The risk factors were further included in the multivariate Logistic regression analysis according to the univariate Logistic regression analysis. After analysis, we obtained: clinical T stage of bladder urothelial carcinoma (OR=0.446, 95%Cl: 0.241-0.826, P=0.01), degree of differentiation (OR=0.501, 95%Cl: 0.262-0.956, P=0.036) ) and CA-IX expression intensity (OR=2.325, 95%Cl: 1.157-4.673, P=0.018) were independent influencing factors for predicting the recurrence of bladder urothelial carcinoma after resection (P<0.05); The number of tumors (OR=0.502, 95%Cl: 0.266-0.947, P=0.033), tumor diameter (OR=1.241, 95%Cl: 0.651-2.364, P=0.512) and were not statistically significant with recurrence after resection (P>0.05). See Table 4 for details.
Table 4: Multivariate Logisti regression analysis of bladder urothelial carcinoma recurrence
parameter
|
OR
|
95% confidence interval for EXP(B)
|
P value
|
lower limit
|
upper limit
|
T stage
|
0.446
|
0.241
|
0.826
|
0.01
|
Differentiation
|
0.501
|
0.262
|
0.956
|
0.036
|
number of tumors
|
0.502
|
0.266
|
0.947
|
0.033
|
tumor diameter
|
1.241
|
0.651
|
2.364
|
0.512
|
CA-IX expression
|
2.325
|
1.157
|
4.673
|
0.018
|
2.5 Kaplan-Meier survival curve of CA-IX expression intensity and recurrence.
2.5.1 This study enrolled 245 patients who underwent transurethral resection of bladder tumors in our hospital, and a total of 194 patients were followed up for 5 years. Urinary tract B-ultrasound and cystoscopy were evaluated every 3 months after surgery. Relapse status, time of relapse and clinical information of patients were recorded. During the follow-up period, 76 cases recurrence, 118 cases did not recured, and the recurrence rate was 39.17% (76/194). Among them, 59 cases relapsed in CA-IX positive expression group, the recurrence rate was 44.69% (59/132), and 17 cases relapsed in the negative expression group, and the recurrence rate was 27.41% (17/62). The mean recurrence time of CA-IX positive group was 29.93±9.86 (months), and the mean recurrence time of CA-IX negative group was 34.02±12.44 (months), P=0.038 by T test of the two observation groups. The results showed that the recurrence rate and recurrence time of patients with positive expression of CA-IX in bladder urothelial carcinoma were significantly higher than those of patients with negative expression of CA-IX.
2.5.2 According to the expression of CA-IX, the relapsed patients were divided two groups, and then the Kaplan-Meier survival curve was drawn. The curve showed that the recurrence rate of CA-IX positive expression group was significantly higher than that of negative group (P<0.05). See Figure D for details.