| Part I The clinical and pathological features of patients with bladder cancer associated with diabetes mellitusObjectives:Analyze and summarize the clinical features and histological features of the tumor of Chinese population of patients with bladder cancer associated with diabetes mellitus (DM).Methods:The clinical information of the patients with bladder cancer which was treated in the Department of Urology, Second Hospital of Tianjin Medical University in January2008-December2011period were collected, a total of992. The patients with bladder cancer were divided into bladder cancer with DM group and bladder cancer without DM group. The clinical and pathological characteristics of the two groups of patients were analyzed.Results:Comparing the two groups, the body weight and BMI was significantly higher in bladder cancer with DM group than bladder cancer without DM group (P=0.000、P=0.000). The level of FPG was significantly higher in bladder cancer with DM group than bladder cancer without DM group (P=0.000). The proportion of patients with hypertension was significantly higher in bladder cancer with DM group than bladder cancer without DM group (P=0.001). Compared with bladder cancer without DM group, the tumors were more occurred in bladder neck (P=0.002), the tumor grade was higher (P=0.042), the proportion of distant metastases was higher (P=0.000) in bladder cancer with DM group. No significant difference was seen between two groups when the gender, smoking, tumor stage, tumor size, tumor numbers and lymph node metastasis were assessed.Conclusion:DM increases the body weight and BMI of patients with bladder cancer. Compared with bladder cancer without DM group, the level of FPG and the proportion of patients with hypertension were significantly higher in bladder cancer with DM group. The tumors were more occurred in bladder neck and the tumor grade was higher in bladder cancer with DM group. The higher proportion of distant metastases in bladder cancer with DM means that DM may increase the degree of malignancy of bladder cancer. Part Ⅱ The expression of GLUT1in tumor tissue in UCC and it correlated research with HIFla and angiogenesisObjectives:By studying the expression of GLUT1in urothelial cell carcinomas of the bladder (UCC), to explore the abnormal expression of GLUT1in UCC with DM, and it correlation with tumor angiogenesis and regulatory mechanisms.Methods:The expression of GLUT1、HIF1a、MVD and VEGF was detected in UCC by immunohistochemistry. Differences in each index were compared between bladder cancer with DM group and bladder cancer without DM group. The correlation between GLUT1and HIF1a、MVD. VEGF was analyzed.Results:GLUT1was not expressed in adjacent normal bladder tissues, but was overexpressed in UCC tissues. And the expression of GLUT1was correlated with tumor staging (P=0.000) and grading (P=0.000). The expression of GLUT1was significantly higher in UCC with DM than UCC without DM (P=0.015). The expression of HIF1a was correlated with tumor staging(P=0.000), grading(P=0.000) and DM (P=0.015). The expression of MVD and VEGF was correlated with tumor staging and grading (P=0.000、P=0.001; P=0.000、P=0.039), but was not correlated with DM(P>0.05; P>0.05). The expression of GLUT1was positively correlated with HIF1a、MVD and VEGF in UCC (r=0.794, P=0.000; r=0.549, P=0.006; r=0.381, P=0.006).Conclusion:The expression of GLUT1was correlated with tumor grading, tumor staging, tumor hypoxic microenvironment and angiogenesis. And DM will increase the expression of GLUT1in tumor tissue in UCC.Part Ⅲ The interference effect of GLUT1shRNA for the bladder cancer cell line T24Objectives:By studying the effect of GLUT1on bladder cancer, To explore the role of GLUT1in progress of bladder cancer.Methods:Through in vitro studies using recombinant plasmid cytology technique to interference GLUT1level in bladder cancer cells, through Q-PCR、western blotting to detect the interference effect of RNA, through MTT, Flow cytometry and transwell to verify the effect of GLUT1for bladder tumor cell proliferation, apoptosis, cycle and invasion. By using2-deoxy-D-[3H]-glucose method to detect the changes in the rate of glucose uptake in cells.Results:By using A549tumor cells, we screened GLUT1-shRNA which the interference effect for the GLUT1was up to82%. The results of MTT, Flow cytometry and transwell shows that GLUT1-shRNA inhibited proliferation of T2458.69%(48h), promote apoptosis and necrosis of T24, the rate48h necrosis and apoptosis was47.41%and22.03%, inhibit the invasion ability of T24cells. GLUT1-shRNA reduces glucose uptake rate was62.39%in the T24cells.Conclusion:GLUT1express in human tumor cells, including human bladder cancer cells. Successfully constructed, screened pYr-1.1-GLUT1-shRNA Plasmid, the interference effects for the GLUT1was up to82%. In vitro GLUT1-shRNA can reduces glucose uptake in the T24bladder cancer cells, inhibition of proliferation, apoptosis, inhibition of invasion effect. GLUT1may play an important role in the progression of bladder cancer.Part Ⅳ Analysis of the polymorphisms of the GLUT1gene in urothelial cell carcinomas of the bladder and its correlation with P53, Ki67and GLUT1expressionsObjectives:The glucose transporter1(GLUT1) is a key rate-limiting factor in the transport and metabolism of glucose in tumor cells. Also recent studies have found that some single nucleotide polymorphisms (SNPs) of the GLUT1gene are associated with kinds of cancers’risk.Methods:Frequencies of two GLUT1SNPs (XbaⅠ G>T and HaeⅢ T>C) were investigated with PCR-RFLP method in314patients with urothelial cell carcinomas of the bladder (UCC) and204normal persons. And the expression of the P53, Ki67and GLUT1were assayed in51paraffin-embedded specimens of UCC patients by immunohistochemistry.Results:For the GLUT1XbaI G>T polymorphism, there was a decrease in the frequency of the TT genotype in the patient population versus the controls (38%vs.53%, P=0.033). This corresponded to an increase in the GT genotype in the patient population versus the controls (58%vs.43%, P=0.034). The T allele was decreased in frequency in the patient population versus the controls (64%vs.74%, P=0.022). No significant difference was seen between the patient population and the control population when the GG genotype was assessed. For the GLUT1HaeⅢ T> C polymorphism, there was, a decrease in the frequency of the CC genotype in the patient population versus the controls (4%vs.19%, P<0.001). There was an increase in the frequency of the TC genotype in the patient population versus the controls (83%vs.73%, P=0.006). There was a decrease in the frequency of the C allele in the patient population versus the controls (45%vs.56%, P=0.001). No significant difference was seen between the patient population and the control population when the TT genotype was assessed. The GLUT1Xba1genotype GG was more frequent in patients with higher tumor stage (P=0.014) and higher tumor grade (P=0.039). The GG genotype of the Xba1G>T SNP was significantly associated with higher Remmele immunoreactive score (IRS) of Ki67(P=0.031) and higher IRS of GLUT1(P=0.009). The Xba1G>T SNP showed no association with IRS of P53(P=0.935). The HaeⅢ T>C SNP showed no association with IRS of P53(P=0.964), IRS of ki67(P=0.145), IRS of GLUT1(P=0.167).Conclusion:The TT genotype of XbaⅠ G>T SNP and CC genotype of HaeⅢ T> C SNP may have protective effect in the carcinogenesis process of UCC. The GG genotype of the XbaⅠ G>T SNP was positively associated with tumor proliferation, glucose metabolism, tumor grade and stage. Therefore, the GLUT1might become a possible proliferation-related prognostic factor for UCC. |
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