| Objective To detect the expression of EZH2, Bmi-1, Survivin and Survivin-3B genes in transitional cell of bladder(TCC), explore their clinical significance, and reveal the effect of their expression.Methods 1.The fresh carcinoma tissues of 70 patients with TCC and 15 normal bladder mucosa from March 2006 to June 2009 were collected. Quantitative real-time reverse transcription and polymerase chain reaction( Real time PCR) was used to analyze the expression of EZH2, Bmi-1, Survivin and Survivin-3B genes in the bladder carcinoma tissue and normal bladder mucosa. The patient group could be divided into three subgroups according to the world Health Organization(WHO) of pathologic grades in 2004: papillary urothelial neoplasms of low malignant potential(PUNLMP) group(5 cases),low-grade group(23 cases) and high-grade group(42 cases) and two subgroups according to the TNM classification of malignant tumors in 2002:36 cases of superficial group(Tis,Ta,T1) and 34 cases of invasive group(T2-T4). 2. Fresh urine of 39 patients with TCC and 13 healthy volunteers who voided the second urine in morning from March 2009 to October 2009 was collected. The expression of EZH2 and Bmi-1 genes in urine was detected by R/Q real time PCR. The patient group could be divided into two subgroups according to the WHO of pathologic grades in 2004: low-grade group(15 cases) and high-grade group(24 cases) and two subgroups according to the TNM classification of malignant tumors in 2002: 22 cases of superficial group(Tis,Ta,T1) and 17 cases of invasive group(T2-T4).Results1. EZH2 gene expressed in tissue and urine of transitional cell of bladder ⑴The relative expression of EZH2 gene was 0.016±0.237 in normal bladder mucosa. In TCC patients, the relative expression of EZH2 gene was 0.145±0.188, there were significant statistical differences compared with the control group, P values was 0.000. Different expression level of EZH2 mRNA could be observed among different pathologic grades, PUNLMP group: 0.108±0.188, low-grade group: 0.074±0.098 and high-grade group: 0.189±0.214. The expression of EZH2 mRNA also differed between clinical stages: superficial group: 0.0868±0.1233 and invasive group: 0.2069±0.2241. The differences between low-grade group, high-grade group and superficial group and invasive group were statistically significant (P<0.05).⑵The relative expression of EZH2 gene was 0.002±0.003 which was very low in healthy volunteers. In the urine of TCC patients, the relative expression of EZH2 gene was 0.038±0.060, there was significant statistical differences compared with the control group, the P values was 0.000. Different expression level of EZH2 mRNA could be observed among different pathologic grades in urine, low-grade group: 0.010±0.016 and high-grade group: 0.056±0.072. The expression of EZH2 mRNA also differed between clinical stages: superficial group: 0.009±0.014 and invasive group: 0.081±0.077. The difference between different pathologic grades and different clinical stages were statistically significant (P<0.05).2. Bmi-1 gene expressed in tissue and urine of transitional cell of bladder⑴The relative expression of Bmi-1 gene was 0.175±0.097 in normal bladder mucosa. In TCC patients, the relative expression of Bmi-1 gene was 0.689±0.793, there were significant statistical differences compared with the control group, the P values was 0.001. Different expression level of Bmi-1 mRNA could be observed among different pathologic grades, PUNLMP group: 0.665±0.767, low-grade group: 0.496±0.455 and high-grade group: 0.799±0.926. The expression of Bmi-1 mRNA also differed between different clinical stages: superficial group: 0.551±0.499 and invasive group: 0.837±1.004.But the difference between different pathologic grades and different clinical stages were not statistically significant (P>005). ⑵The relative expression of Bmi-1 gene was 0.030±0.037 in healthy volunteers. In the urine of TCC patients, the relative expression of Bmi-1 gene was 0.073±0.079, there was significant statistical differences compared with the control group, the P values was 0.029. Different expression level of Bmi-1 mRNA could be observed between different pathologic grades and different clinical stages in urine, low-grade group: 0.044±0.045, high-grade group: 0.091±0.167, superficial group: 0.041±0.039 and invasive group: 0.072±0.076. But the difference between different pathologic grades and different clinical stages were not statistically significant (P>005).3. Survivin and Survivin-3B genes expressed in tissue of transitional cell of bladder⑴The relative expression of Survivin and Survivin-3B genes were 0.241±0.209 and 0.006±0.007 in normal bladder mucosa. In TCC patients, the relative expression of Survivin and Survivin-3B genes were 1.857±3.239 and 0.199±0.447, there were statistically significant compared with the control group, the P values was 0.003 and 0.000.⑵Different expression level of Survivin mRNA could be observed among different pathologic grades, PUNLMP group: 0.267±0.337, low-grade group: 0.856±1.122 and high-grade group: 2.609±3.938. The difference was not statistically significant (P>0.05) between PUNLMP group and low-grade group. The expression of Survivin mRNA also differed between clinical stages: superficial group: 0.873±1.270 and invasive group: 2.899±4.248. But the differences between low-grade group, high-grade group and superficial group and invasive group were not statistically significant (P<0.05).⑶Different expression level of Survivin-3B mRNA could be observed among different pathologic grades, PUNLMP group: 0.019±0.025, low-grade group: 0.096±0.129 and high-grade group: 0.277±0.556. The expression of Survivin-3B mRNA also differed between clinical stages: superficial group: 0.108±0.216 and invasive group: 0.295±0.589. The difference between different pathological grade and different clinical stages were statistically significant (P<0.05).Conclusion 1. The expression of EZH2 gene in tissue and urine of normal people detected scarcely.The expression of EZH2 gene in tissue and urine of TCC was highly related to pathologic grades and clinical stages. The more expression of EZH2 gene,the worse clinical-pathologic feature and higher invasion. The expression of EZH2 gene in urine cells of TCC was paralleled with that in tissue of TCC. So,the oncogenesis of TCC may be regulated by the dysfunction of EZH2 gene transcription in the level of cancer stem cells.2.Survivin gene could be detected in TCC and normal bladder mucosa, the relative expression of that in TCC was higher than the normal bladder mucosa group(P<0.05). The more expression of Survivin, the worse clinical-pathologic features and higher invasion.The relative expression of Survivin-3B gene was low in normal bladder mucosa, and increased with the increase of pathological grade and clinical stage. So, Survivin-3B and Survivin genes play an important role in the occurrence and progression of TCC.The Survivin-3B gene, in Survivin splice variants family, may be the most important target gene in inhibition of TCC apoptosis. 3. In TCC, EZH2, Bmi-1, Survivin and Survivin-3B genes were over-expressed, Which had unfavourable prognosis. 4. The expression levels of EZH2 and Survivin-3B gene in tissues and urine of TCC could have potential values to appraisal tumor progression and recurrence rate of TCC.
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