OBJECTIVETo detect the expressions of receptor tyrosine kinases (RTKs) mRNA and protein and to explore potentially promising tumor markers and conceivable drug target in bladder cancer.

METHODSThe expressions of RTKs mRNA and protein in tissue from invasive urothelial carcinoma of the bladder were examined by real-time quantitative PCR array and cytokine antibody array, with normal bladder tissue as control. The Results were analyzed using bioinformatic approaches.

RESULTSThe expressions of TGFA, STAB1, SERPINE1, ANGPT2, SPINK5, ANGPTL1, PROK1, MDK, CXCL9, GRN, RUNX1, VEGFA, and TGFB1 were obviously upregulated in bladder cancer tissue, while those of EDIL3, PTN, CCL2, PDGFD, FGF13, KITLG, FGF2, SERPINF1, and TNF were downregulated. ALK, Btk, EphB2, ErbB4, PDGFR-α, ROS, Tie-2, Tyk2, and VEGFR3 were over-expressed in bladder cancer, while FRK, Fyn, IGF-IR, Insulin R, Itk, JAK1, JAK3, and LCK were low-expressed.

CONCLUSIONVascular endothelial growth factor/platelet-derived growth factor-targeted therapies may play an active role in treating carcinoma of bladder.

Carcinoma, Transitional Cell ; metabolism ; Humans ; RNA, Messenger ; genetics ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism ; Urinary Bladder Neoplasms ; metabolism

The expression of survivin, a member of inhibitor of apoptosis (IAP) family, was examined in bladder transitional cell cancer (BTCC) tissue and adjacent normal tissues to examine its clinical implication in the development of BTCC. Thirty specimens of bladder cancer were detected for the expression of survivin by using immunohistochemistry and real-time quantitative reverse transcription polymerase chain reaction (RT-QPCR) in BTCC tissue and adjacent normal tissues. Our results showed that the positive rate of survivin immunostaining specimen were 0 and 60% (18/30) in the adjacent normal tissues, bladder cancer, respectively. The-DeltaDeltaCT value of survivin in bladder cancer tissue was 10.2829 (9.0034-11.5624) times that in the adjacent normal tissues. The expressions of survivin were correlated with the pathological grades of tumor and clinical stages. It is concluded that there was only weak expression of survivin mRNA in the adjacent normal tissues, but the expression of survivin mRNA in bladder cancer tissue was much higher than that in the adjacent normal tissues and the expression of survivin was correlated with pathological grades and clinical stages of tumor.
*Carcinoma, Transitional Cell/metabolism ; *Carcinoma, Transitional Cell/pathology ; Microtubule-Associated Proteins/genetics ; Microtubule-Associated Proteins/*metabolism ; Prognosis ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Tumor Markers, Biological/genetics ; Tumor Markers, Biological/*metabolism ; Urinary Bladder Neoplasms/*metabolism ; Urinary Bladder Neoplasms/*pathology

OBJECTIVETo investigate the mRNA expression of platelet-derived endothelial cell growth factor (PD-ECGF) in superficial bladder cancer and its significance.

METHODSPD-ECGF mRNA expressions were determined by RT-PCR in 28 cases of superficial bladder cancers and 6 cases of normal bladder mucosa. The relation between PD-ECGF mRNA expression and tumor invasion to lamina propria or recurrence after transurethral resection was also analyzed.

RESULTSSome degree of PD-ECGF mRNA expression was present in all the samples. The PD-ECGF mRNA level was 3.1-fold higher in pT(1) tumors than in normal bladder mucosa (t = 2.13, P < 0.05) and 2.2-fold higher in pT(1) tumors than in pT(a) tumors (t = 2.66, P < 0.05); G(3) tumors expressed 3.3-fold higher PD-ECGF mRNA than normal bladder mucosa (t = 2.44, P < 0.05) and 2.5-fold higher than G(1 - 2) tumors (t = 3.36, P < 0.01). Eleven cases recurred during the mean follow-up period of 18 months. Three-fold higher PD-ECGF mRNA expression was showed in cases who recurred after transurethral resection than that in cases who did not recur (t = 4.49, P < 0.01). The specificity and sensitivity of predicting tumor recurrence were 82.4% and 81.8% respectively using 0.095 as a cutoff value of PD-ECGF mRNA level in this group of superficial bladder cancer.

CONCLUSIONPD-ECGF mRNA expression correlates with tumor dedifferentiation and plays an important role in the early invasion in superficial bladder cancer. To analyze the PD-ECGF mRNA level contributes to the evaluations of tumor differentiation and invasion to lamina propria as well as recurrence prediction in superficial bladder cancer.

Carcinoma, Transitional Cell ; metabolism ; pathology ; Follow-Up Studies ; Humans ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Thymidine Phosphorylase ; genetics ; metabolism ; Urinary Bladder Neoplasms ; metabolism ; pathology

OBJECTIVETo investigate the relationship between mRNA expression of platelet-derived endothelial cell growth factor (PD-ECGF) and invasion of bladder transitional cell carcinoma (BTCC).

METHODSThe mRNA expression of PD-ECGF in BTCC was detected by RT-PCR. The target PCR bands were analyzed by NIH Image 1.62 software.

RESULTSThe mRNA level of PD-ECGF in BTCC was 3.86 times as high as that of normal bladder mucosa (t = 2.36, P < 0.05). The expression level of stage Ta, T1 and T2-4 tumor was 1.33, 4.02 and 7.59 times as high as that of normal bladder mucosa, respectively. That of Grade 3 tumor was 2.27 times as high as that of Grade 1 - 2 tumor (t = 3.52, P < 0.01).

CONCLUSIONThe mRNA expression of PD-ECGF was positively correlated with the invasiveness and grade of BTCC. The results suggest that the mRNA level of PD-ECGF might be used as an indicator of tumor progression and a guide for clinical treatment of bladder transitional cell carcinoma.

Adult ; Aged ; Carcinoma, Transitional Cell ; metabolism ; pathology ; Cell Differentiation ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; RNA, Messenger ; analysis ; Thymidine Phosphorylase ; genetics ; Urinary Bladder Neoplasms ; metabolism ; pathology

OBJECTIVETo examine the subcellular location of UCA1, a non-coding RNA, analyze its tissue expression pattern, and investigate the relationship between UCA1 expression and bladder carcinoma progression.

METHODSElectron microscopic in situ hybridization technique was employed to determine the subcellular localization of UCA1 gene. RT-PCR was used to detect its mRNA expression level in various tissues.

RESULTSElectron microscopy identified scattered colloidal gold particles on the cell membrane and massive homogeneously distributed particles in the cytoplasm without specific aggregation in the cells; scattered particles were also detected in the cell nuclei. UCA1 gene was overexpressed in the chorionic villi, placenta and fetal bladder tissues. In adult human tissues, UCA1 gene was not expressed except in the heart and spleen. The expression level of UCA1 was increased in 8 common tumor tissues as compared with that in the corresponding normal tissues. UCA1 mRNA was not detected in normal bladder, normal kidney, renal cancer or hyperplastic prostate tissues, but highly expressed in cancerous bladder tissues.

CONCLUSIONUCA1 gene locates in the cytoplasm, and its mRNA expression level is closely correlated to the progression of bladder cancer, indication its potential as a specific molecular marker of bladder cancer.

Aged ; Aged, 80 and over ; Biomarkers, Tumor ; genetics ; metabolism ; Carcinoma, Transitional Cell ; metabolism ; Cell Line, Tumor ; Cells, Cultured ; Cytoplasm ; metabolism ; Female ; Humans ; Male ; Middle Aged ; RNA, Long Noncoding ; RNA, Untranslated ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Urinary Bladder Neoplasms ; metabolism

Objective: To investigate the effect of long non-coding RNA urothelial carcinoma-associated 1 (UCA1) gene on the proliferation, migration, apoptosis and immune escape of endometrial cancer cells and its molecular mechanism. Methods: Endometrial cancer tissues and adjacent normal tissues of patients with endometrioid adenocarcinoma who underwent total or partial hysterectomy in Henan Provincial People's Hospital from 2017 to 2019 were collected. The expressions of UCA1 and miR-204-5p were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), and the cell proliferation, migration and apoptosis were detected by cell counting kit 8 (CCK8) method, Transwell method, flow cytometry, and dual-luciferase reporter assay was used to explore the target relationship between UCA1 and miR-204-5p. HEC-1A-sh-NC or HEC-1A-sh-UCA1 cells were co-cultured with peripheral blood mononuclear cells (PBMC) or cytokine-induced killer cells in vitro to explore the role of UCA1 in immune escape. Results: The expression level of UCA1 in endometrial cancer tissue (17.08±0.84) was higher than that in adjacent normal endometrial tissue (3.00±0.37), and the expression level of miR-204-5p (0.98±0.16) was lower than that in adjacent normal endometrial tissue (2.00±0.20, P<0.05). Pearson correlation analysis showed that the expression of miR-204-5p was negatively correlated with the expression of UCA1 (r=-0.330, P=0.030). The expressions of UCA1 and miR-204-5p were associated with the International Federation of Gynecology and Obstetrics stage of endometrial cancer, lymph node metastasis and vascular invasion (P<0.05). The relative ratio of absorbance (0.58±0.11) and the number of cell migration [(199.68±18.44)] in the sh-UCA1 group were lower than those in the sh-NC group (1.24±0.17 and 374.76±24.83), respectively. The apoptosis rate of sh-UCA1 group [(28.64±7.80)%] was higher than that of sh-NC group [(14.27±4.38)%, P<0.05]. After different ratios of effector cells and target cells were cultured, the cell survival rate of HEC-1A-sh-UCA1 group was lower than that of HEC-1A-sh-NC group, and the difference was statistically significant (P<0.05). UCA1 had a binding site for miR-204-5p. The relative ratio of absorbance (1.74±0.08) and the number of cell migration (426.00±18.00) cells in the UCA1+ anti-miR-204-5p group were higher than those in the control group [1.00±0.03 and (284.00±8.00) cells, respectively]. The apoptosis rate of UCA1+ anti-miR-204-5p group [(5.42±0.93)%] was lower than that of control group [(14.82±1.48)%, P<0.05]. HEC-1A-sh-UCA1 cells could induce higher interferon gamma (IFN-γ) expression when co-cultured with PBMC, and the levels of IFN-γ expression in PHA group and PHA+ pre-miR-204-5p group cells were 2.42±0.49 and 1.88±0.26, which were higher than that in the PHA+ pre-NC group (0.85±0.10, P<0.05). When co-cultured with cytokine-induced killer cells (different ratios) in vitro, the HEC-1A-sh-UCA1 group and the HEC-1A-pre-miR-204-5p group had lower survival rates than that in the HEC-1A-pre-miR-204-5p group. In the HEC-1A-pre-NC group, the differences were statistically significant (P<0.05). Conclusion: UCA1/miR-204-5p may play an important role in human endometrial cancer.
Female ; Humans ; MicroRNAs/metabolism* ; RNA, Long Noncoding/genetics* ; Leukocytes, Mononuclear ; Carcinoma, Transitional Cell ; Antagomirs ; Cell Line, Tumor ; Urinary Bladder Neoplasms ; Cell Proliferation ; Endometrial Neoplasms/genetics* ; Apoptosis/genetics* ; Cell Movement/genetics* ; Gene Expression Regulation, Neoplastic

PURPOSE: Our previous high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry study identified bladder cancer (BCA)-specific urine metabolites, including carnitine, acylcarnitines, and melatonin. The objective of the current study was to determine which metabolic pathways are perturbed in BCA, based on our previously identified urinary metabolome. MATERIALS AND METHODS: A total of 135 primary BCA samples and 26 control tissue samples from healthy volunteers were analyzed. The association between specific urinary metabolites and their related encoding genes was analyzed. RESULTS: Significant alterations in the carnitine-acylcarnitine and tryptophan metabolic pathways were detected in urine specimens from BCA patients compared to those of healthy controls. The expression of eight genes involved in the carnitine-acylcarnitine metabolic pathway (CPT1A, CPT1B, CPT1C, CPT2, SLC25A20, and CRAT) or tryptophan metabolism (TPH1 and IDO1) was assessed by RT-PCR in our BCA cohort (n=135). CPT1B, CPT1C, SLC25A20, CRAT, TPH1, and IOD1 were significantly downregulated in tumor tissues compared to normal bladder tissues (p<0.05 all) of patients with non-muscle invasive BCA, whereas CPT1B, CPT1C, CRAT, and TPH1 were downregulated in those with muscle invasive BCA (p<0.05), with no changes in IDO1 expression. CONCLUSION: Alterations in the expression of genes associated with the carnitine-acylcarnitine and tryptophan metabolic pathways, which were the most perturbed pathways in BCA, were determined.
Aged ; Biomarkers/metabolism ; Carcinoma, Transitional Cell/genetics/*metabolism/pathology ; Carnitine/*analogs & derivatives/genetics/metabolism ; Case-Control Studies ; Female ; Humans ; Male ; Metabolic Networks and Pathways/*physiology ; Middle Aged ; RNA, Messenger/metabolism ; Real-Time Polymerase Chain Reaction ; Urinary Bladder Neoplasms/genetics/*metabolism/pathology

OBJECTIVETo investigate the differential expression of the hyaluronic acid synthase (HAS) family in human bladder transitional cell carcinoma (BTCC) and its potential clinical significance.

METHODSThe relative quantitative detection of the expression of HAS isoforms (HASs) was performed in 78 human BTCC tissues (mRNA & protein) and 12 normal human bladder mucosa (protein) by real-time RT-PCR and Western blot, and the results were statistically analyzed according to the clinical data.

RESULTSAll the BTCC tissues expressed three HAS isoform mRNA and protein, but to a different extent, as for mRNA, HAS3 > HAS2 > HAS1 (P < 0.001), with a significant difference in HAS1/HAS2, HAS1/HAS3 and HAS2/HAS3 (P = 0.003, < 0.001, 0.006, respectively). Among the proteins, the HAS2 expression was the highest, with a significant difference in HAS1/HAS2, and HAS2/HAS3 (P = 0.004, 0.001, respectively), but not in HAS1/HAS3. The elevation of HAS1 mRNA and protein expression was significantly related with the tumor malignancy, tumor initial onset/recurrence, T1/T2 and T1/T3-4 stags, and tumor grading (P = 0.02, < 0.001, 0.038, < 0.001; 0.025, 0.031, 0.023, 0.002; respectively). The HAS2 mRNA expression was significantly related with tumor size (diameter ≤ 3.0 cm/> 3.0 cm), tumor number (single or multiple), tumor initial onset/recurrence, T-staging, and histopathological differentiation (low grade/high grade) (P = 0.012, 0.004, < 0.001, < 0.001, < 0.001, respectively), but its protein expression was not significantly different in all subgroups except with the tumor size (mass diameter > 3.0 cm/≤ 3.0 cm). However, HAS3 mRNA and protein expression had no significant difference among all the subgroups. In normal human bladder mucosa, no HAS expressions were detected.

CONCLUSIONSThe abnormally high expression of the HASs further indicate the reliability of hyaluronan as a urinary marker for human BTCC. Compared with HAS1 and HAS3, HAS2 as a marker may have more usefulness in studies on human BTCC carcinogenesis or development. The high expression of HAS1 protein seems to play a more important role in the BTCC tumorigenesis, and may indicate a poor prognosis of the BTCC patients.

Biomarkers, Tumor ; Blotting, Western ; Carcinoma, Transitional Cell ; genetics ; metabolism ; Glucuronosyltransferase ; metabolism ; Humans ; Hyaluronan Synthases ; Hyaluronic Acid ; metabolism ; Neoplasm Recurrence, Local ; RNA, Messenger ; biosynthesis ; Reproducibility of Results ; Urinary Bladder Neoplasms ; genetics ; metabolism

OBJECTIVETo determine the levels of MMP-2 and COX-2 mRNA in bladder transitional cell carcinoma tissues and explore their relationship.

METHODSWe enrolled in this study 42 patients with bladder transitional cell carcinoma, including Ta-T1 (n = 18), T2-T4 (n = 24), G1 (n = 12), G2 (n = 19), G3 (n = 11), metastasis (n =26) and non-metastasis (n = 16). Another 5 cases of normal bladder tissues were taken as controls, and the levels of MMP-2 and COX-2 mRNA were detected by RT-PCR.

RESULTSThe relative expressions of COX-2 mRNA were 1.038 +/- 0. 484 in Ta-T1, 1.489 +/- 0.584 in T2-T4, 0.920 +/- 0.442 in G1, 1.338 +/- 0.584 in G2 and 1.632 +/- 0.515 in G3, all significantly higher than that of the controls (0.460 +/- 0.224, P < 0.05). And the corresponding relative levels of MMP-2 mRNA were 1.107 +/- 0.384, 1.604 +/- 0.425, 0.971 +/- 0.370, 1.445 +/- 0.378 and 1.755 +/- 0.387, also significantly higher than that of the latter group (0.423 +/- 0.227, P < 0.05). The COX-2 and MMP-2 mRNA levels in the tumor tissues with and without metastasis were 1.591 +/- 0.455 vs 0.815 +/- 0.430 and 1.676 +/- 0.339 vs 0.927 +/- 0.228, (P < 0.01), respectively, with a positive correlation between the mRNA level of COX-2 and that of MMP-2 (r = 0. 703, P < 0.01).

CONCLUSIONMMP-2 and COX-2 mRNA are highly expressed in bladder transitional cell carcinoma tissues and their expressions are positively correlated with the degree of malignancy. MMP-2 and COX-2 might play a synergetic role in the pathogenesis and progression of bladder transitional cell carcinoma.

Adult ; Aged ; Carcinoma, Transitional Cell ; metabolism ; pathology ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Female ; Humans ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; genetics ; Urinary Bladder Neoplasms ; metabolism ; pathology

OBJECTIVETo evaluate the anticancer effects of exogenous human WT-PTEN overexpression on bladder transitional carcinoma cell line EJ.

METHODSThe plasmid containing WT-PTEN or mutant PTEN was separately transfected into bladder transitional carcinoma cell line EJ, and the protein expression of PTEN in the EJ cells was detected by Western blot. Cell morphological changes were observed under the inverted microscope and transmission electron microscope. MTT test was used to assess the effect of PTEN on proliferation and anticancer effects for mitomycin and theraubicin. The change of bcl-2 expression in the cells was measured by Western blot. The empty plasmid was used as control.

RESULTSWestern blot analysis showed that EJ cells expressed high level of PTEN protein after transfection with WT-PTEN or mutant PTEN plasmid. Abnormal morphological changes of the cells were observed in WT-PTEN transfected groups. The growth of EJ cells treated with WT-PTEN was significantly inhibited by 40.1% and anticancer effects were enhanced by mitomycin and theraubicin, but the cells transfected with mutant PTEN plasmid did not show such similar biological behavior.

CONCLUSIONWT-PTEN gene transfection can suppress the in vitro growth and induce apoptosis of bladder transitional carcinoma cell line EJ cells. Mutant PTEN does not show similar biological behavior. Overexpression of WT-PTEN inhibits cancer cell proliferation by down-regulating bcl-2 expression in the cells.

Antibiotics, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Carcinoma, Transitional Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Doxorubicin ; analogs & derivatives ; pharmacology ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Electron, Transmission ; Mitomycin ; pharmacology ; Mutation ; PTEN Phosphohydrolase ; genetics ; metabolism ; physiology ; Plasmids ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; physiology ; Transfection ; Urinary Bladder Neoplasms ; genetics ; metabolism ; pathology