OBJECTIVES: To investigate the role of apelin in regulating proliferation, migration and angiogenesis of bladder cancer cells and the possible regulatory mechanism. METHODS: GEO database was used to screen the differentially expressed genes in bladder cancer tissues and cells. Bladder cancer and paired adjacent tissues were collected from 60 patients for analysis of apelin expressions in relation to clinicopathological parameters. In cultured bladder cancer J82 cells and human umbilical vein endothelial cells (HUVECs), the effects of transfection with an apelin-overexpressing plasmid or specific siRNAs targeting apelin, fibroblast growth factor 2 (FGF2) and fibroblast growth factor receptor 1 (FGFR1) on proliferation and migration of J82 cells and tube formation in HUVECs were examined using plate cloning assay, Transwell assay, and angiogenesis assay; the changes in FGF2 expression and FGFR1 phosphorylation were detected using Western blotting. RESULTS: The expression level of apelin was significantly higher in bladder cancer tissues than adjacent tissues, and bladder cancer cell lines (T24 and J82) also expressed higher mRNA and protein levels of apelin than SV-HUC-1 cells. Apelin expression level in bladder cancer tissues was correlated with tumor invasion, distant metastasis and advanced TNM stages. Apelin knockdown significantly suppressed proliferation and migration of J82 cells and decreased the total angiogenic length of HUVECs. In contrast, apelin overexpression significantly promoted proliferation and migration and enhanced FGFR1 phosphorylation in J82 cells, and increased the total angiogenesis length in HUVECs, but this effects were effectively mitigated by transfection of the cells with FGF2 siRNA or FGFR1 siRNA. CONCLUSIONS High expression of apelin promotes J82 cell proliferation and migration and HUVEC angiogenesis by promoting activation of the FGF2/FGFR1 pathway.
Humans ; Urinary Bladder Neoplasms/blood supply* ; Receptor, Fibroblast Growth Factor, Type 1/metabolism* ; Cell Proliferation ; Cell Movement ; Fibroblast Growth Factor 2/metabolism* ; Neovascularization, Pathologic ; Human Umbilical Vein Endothelial Cells ; Cell Line, Tumor ; Signal Transduction ; Apelin ; Intercellular Signaling Peptides and Proteins/genetics* ; Female ; Male ; Angiogenesis

OBJECTIVETo explore the expressions of CD133 and CD82/KAI1 in bladder urothelial carcinoma, their association with the clinicopathological factors and their roles in vasculogenic mimicry (VM) in the tumor.

METHODSThe expressions of CD133 and CD82/KAI1 and VM were detected by immunohistochemistry and histochemistry in 90 specimens of bladder urothelial carcinoma and 20 specimens of normal bladder epithelium tissue.

RESULTSThe positivity rates of CD133, CD82/KAI1 and VM in normal bladder epithelium tissue were 0, 90% and 0, showing significant differences from the rates of 65.6%, 31.1% and 31.1% in urothelial carcinoma, respectively (P<0.01). Positive expressions of CD133, CD82/KAI1 and VM were significantly correlated with pTNM stage and tumor relapse (P<0.01) but not with gender, age, or tumor numbers (P>0.05). CD133 expression was positively correlated with VM (P=0.487, P<0.05), and CD82/KAI1 expression was negatively correlated with VM (r=-0.452, P<0.01) and CD133 (r=-0.776, P<0.05).

CONCLUSIONThe expressions of CD133 and CD82/KAI1 proteins are involved in the occurrence of VM in bladder urothelial carcinoma to contribute to the invasion and relapse of bladder carcinoma.

AC133 Antigen ; Aged ; Aged, 80 and over ; Antigens, CD ; metabolism ; Carcinoma ; blood supply ; metabolism ; Case-Control Studies ; Female ; Glycoproteins ; metabolism ; Humans ; Immunohistochemistry ; Kangai-1 Protein ; metabolism ; Male ; Middle Aged ; Peptides ; metabolism ; Urinary Bladder Neoplasms ; blood supply ; metabolism

OBJECTIVETo study the relationship between enhanced degree in the cancer lesion and angiogenesis in the tumor through early manifestations in enhanced helical CT in patients with bladder cancer.

METHODSFifty-three patients with bladder carcinoma were examined by pelvic plain CT and helical CT scan at peak enhancement of cancer lesion. Histologic grade, vascular endothelial growth factor (VEGF) and micro-vascular density (MVD) were analyzed for each resected cancer lesion. Pearson and Spearman correlation tests were used to assess the relationship between CT enhancement and histologic grade, VEGF or MVD.

RESULTSIn early enhancing phase of helical CT, different degrees of enhancement were observed in 73 bladder cancer lesions. The difference between average CT attenuation and MVD in different histologic grade cancer lesions was statistically significant (P < 0.001). A positive correlation was found between the CT enhancement and MVD (gamma = 0.936, P < 0.001), histologic grade (gamma = 0.75, P < 0.001), but VEGF of bladder cancer did not correlate with the CT enhancement or MVD.

CONCLUSIONThe early enhancement of helical CT enhancement of bladder cancer, showing a positive relation to MVD and histologic grade, can reflect the tumor angiogenesis and blood supply.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Radiographic Image Enhancement ; Tomography, Spiral Computed ; Urinary Bladder Neoplasms ; blood supply ; diagnostic imaging ; pathology ; Vascular Endothelial Growth Factor A ; analysis

OBJECTIVETo verify whether the extracellular domain of kinase domain region (KDR) has anti-angiogenesis activity in vivo.

METHODScDNA was cloned into adeno-associated virus (AAV) vector pSNAV and transfected to baby hamster kidney (BHK) cells. Recombinant AAV was obtained from the cell culture supernatant after adding helper virus. Recombinant AAV-infected human bladder cancer EJ cell line (EJ cells) were injected subcutaneously into Balb-c nude mice. Tumor specimens were removed from the mice, paraffin-embedded and sliced, then stained by immunohistochemistry. Microvessel density (MVD) was determined under a microscope.

RESULTSThe tumor volume developed by EJ cells transfected with the extracellular domain of KDR was significantly smaller (1.70 +/- 0.18 cm(3)) compared with that in the control (5.62 +/- 0.67 cm(3)) (P < 0.05), although tumor developed to be detectable on almost the same time (14.7 +/- 2.4 days vs 14.1 +/- 3.2 days). Further, MVD in the experimental group was lower than that in the control (41.3 +/- 4.8 vs 6.2 +/- 2.1, P < 0.05).

CONCLUSIONThe extracellular domain of KDR could be expressed in nude mouse bladder cancer tissue and inhibit tumor angiogenesis.

Animals ; Cloning, Molecular ; Cricetinae ; Dependovirus ; genetics ; Endothelial Growth Factors ; metabolism ; Female ; Genetic Therapy ; Intercellular Signaling Peptides and Proteins ; metabolism ; Lymphokines ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neovascularization, Pathologic ; prevention & control ; Urinary Bladder Neoplasms ; blood supply ; therapy ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; Vascular Endothelial Growth Factors