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

PURPOSE: To compare the expression of survivin and its association with clinicopathological criteria in major types of urinary bladder carcinoma, specifically, transitional cell carcinoma with and without squamous differentiation and squamous cell carcinoma. MATERIALS AND METHODS: Immunohistochemical staining for survivin and Ki67 was performed on paraffin-embedded sections of 104 carcinomas: 52 transitional cell carcinoma, 20 transitional cell carcinoma with squamous differentiation, and 32 squamous cell carcinoma. Expression of survivin in >10% of tumor cells was described as altered survivin status. Ki67 staining in >20% of tumor cells was described as a high proliferation index. RESULTS: Altered survivin expression was detected in 60/104 specimens (58%) and was significantly more frequent in transitional cell carcinoma (78%) than in squamous cell carcinoma (38%) or transitional cell carcinoma with squamous differentiation (40%) (p<0.0001). In transitional cell carcinoma but not in squamous cell carcinoma, altered survivin status was associated with higher tumor grade, higher proliferation index, and recurrence. In the whole specimens, altered survivin expression was significantly associated with advanced stage (p<0.001), recurrence (p=0.005), distant metastasis (p<0.001), and death (p=0.001). In the multivariate analysis, altered survivin was an independent poor prognostic factor for recurrence. CONCLUSIONS: Unlike in transitional cell carcinoma, alteration of survivin expression in squamous cell carcinoma occurs less frequently and is not associated with features of tumor aggression or patient outcome. These findings raise a question: are urinary bladder carcinoma patients with squamous cell carcinoma type suitable candidates for survivin vaccine? This is an important question to be answered before approving the vaccine in management.
Carcinoma, Squamous Cell/*genetics ; Carcinoma, Transitional Cell/*genetics ; Female ; Humans ; Inhibitor of Apoptosis Proteins/genetics/*metabolism ; Ki-67 Antigen/metabolism ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Grading ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Prognosis ; Treatment Outcome ; Tumor Markers, Biological ; Urinary Bladder/pathology ; Urinary Bladder Neoplasms/*genetics

OBJECTIVETo investigate the changes of gene expression file in transitional cell carcinoma of bladder after hepatocyte cell adhesion molecule(hepaCAM) overexpression.

METHODSAffymetrix Human Genome U133 Plus 2.0 Array was used to investigate the changes of gene expression profile between adenovirus-green fluorescent protein(GFP) -hepaCAM group and GFP group in transitional cell carcinoma of bladder EJ cells.Significant Analysis of Microarray(SAM) was used to screen the differentially expressed genes, DAVID software was used to conduct gene ontology analysis and wikiPathway analysis based on the differentially expressed genes. Reverse transcription-polymerase chain reaction and Western blot were applied to verify microarray data.

RESULTSCompared with the GFP group, a total of 2469 genes were up-regulated or down-regulated by more than 2 times in the GFP-hepaCAM group. Among these genes, 1602 genes were up-regulated and 867 were down-regulated.Most of the differentially expressed genes were involved in the function of cell proliferation and cell cycle regulation. The mRNA expressions of nibrin, liver kinase B1, and cyclin D1 detected by reverse transcription-polymerase chain reaction in three different bladder cancer cell lines were consistent with the microarray data.The protein expressions of nibrin and liver kinase B1 in these three cell lines measured by Western blot were consistent with the mRNA expression.

CONCLUSIONSHepaCAM can alter the gene expression profile of bladder cancer EJ cells. The well-known anti-tumor effect of hepaCAM may be mediated by regulating the gene expression via multiple pathways.

Carcinoma, Transitional Cell ; genetics ; pathology ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cyclin D1 ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; physiology ; Humans ; Nuclear Proteins ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; Proteins ; genetics ; physiology ; Urinary Bladder Neoplasms ; genetics ; pathology

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

Biomarkers, Tumor ; genetics ; metabolism ; Carcinoma, Transitional Cell ; genetics ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Humans ; Ki-67 Antigen ; metabolism ; Microsatellite Repeats ; Mutation ; Neoplasm Grading ; Neoplasm Staging ; Oligonucleotide Array Sequence Analysis ; Pathology, Molecular ; Receptor, Fibroblast Growth Factor, Type 3 ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Urinary Bladder Neoplasms ; genetics ; metabolism ; pathology

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Basigin ; analysis ; Biomarkers, Tumor ; analysis ; genetics ; Carcinoma, Transitional Cell ; drug therapy ; genetics ; metabolism ; Gene Expression Profiling ; Humans ; In Situ Hybridization, Fluorescence ; Inhibitor of Apoptosis Proteins ; analysis ; Mutation ; Neoplasm Recurrence, Local ; metabolism ; Nuclear Proteins ; analysis ; Receptor, Fibroblast Growth Factor, Type 3 ; analysis ; genetics ; Tumor Suppressor Protein p53 ; analysis ; genetics ; Urinary Bladder Neoplasms ; drug therapy ; genetics ; metabolism

OBJECTIVETo study the clinicopathologic features of urothelial hyperplastic lesion with an endophytic growth pattern and the role of immunohistochemistry and multitargeted fluorescence in situ hybridization (FISH) in the differential diagnosis.

METHODSForty-one cases of urothelial lesions exhibiting endophytic growth patterns were reviewed and reclassified as inverted papilloma, urothelial carcinoma with an endophytic growth pattern, and florid von Brunn nest. The gains of chromosomes 3, 7, and 17 and loss of 9p21 was detected by FISH, and performed immunohistochemical staining for CK20, p53, and Ki-67. Follow-up data of 12 cases were obtained.

RESULTS(1) Twelve inverted papillomas sized 1.2 cm in average, consisted of anastomosing cords and nests with uniform width distribution involving the lamina propria, the central portion contained streaming cells with squamous metaplasia, and the periphery showed palisading. No or rare atypia and mitosis were found. Focal exophytic papillary component lined by less than 6 layers of normal urothelium were observed in 4 cases. (2) Twenty-four urothelial carcinomas with an endophytic growth pattern sized 2.1 cm in average, demonstrated the similar architecture with inverted papilloma, but exhibited thick columns and variable thickness of the cords, irregular size and shape of large nests with transition into solids. Mild to moderate cytologic atypia was shown, and mitotic figures ranged 1 to 8 per 10 HPFs. Exophytic papillary component was not observed in 3 cases, but the superficial urothelium showed dysplasia, while coexisted exophytic component in other cases was associated with low malignant potential or low grade tumor. (3) Five florid von Brunn nests sized 0.9 cm in average, had normal or hyperplastic urothelium, variable nests with cysts compacted in lamina propria, no cytologic atypia and mitosis. Twenty-one of 24 (79.1%) urothelial carcinomas with an endophytic growth pattern displayed abnormally positive results by multitargeted FISH, whereas all inverted papillomas and florid von Brunn nests were negative. Immunohistochemically, CK20 was weakly positive in 2 cases of urothelial carcinoma with an endophytic growth pattern, and negative in all inverted papillomas and florid von Brunn nests. p53 weakly stained 5% to 50% nuclei of the tumor cells in 16 cases of urothelial carcinomas with an endophytic growth pattern and 1 inverted papilloma. 1%-5% tumor cells expressed Ki-67 in urothelial carcinoma with an endophytic growth pattern, and less than 1% in inverted papilloma and florid von Brunn nests. Follow-up study revealed that 2 cases of urothelial carcinoma with an endophytic growth pattern had developed invasive carcinoma, underwent cystectomy, and metastasized remotely. No recurrence occurred in cases of inverted papilloma.

CONCLUSIONSBenign and malignant urothelial lesions with an endophytic growth pattern present histologic overlapping. Urothelial carcinoma with an endophytic growth pattern displays unique characteristics in morphology and immunohistochemistry. Multitargeted FISH analysis is helpful in the differential diagnosis.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Transitional Cell ; genetics ; metabolism ; pathology ; surgery ; Chromosome Aberrations ; Diagnosis, Differential ; Follow-Up Studies ; Humans ; Hyperplasia ; In Situ Hybridization, Fluorescence ; Keratin-20 ; metabolism ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Papilloma, Inverted ; genetics ; metabolism ; pathology ; surgery ; Tumor Suppressor Protein p53 ; metabolism ; Urinary Bladder Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Urothelium ; 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 prepare IL-2-anchored and tumor-derived exosomes vaccine, and investigate the antitumor efficiency of the special cytotoxic T-lymphocytes induced by Ex/GPI-IL-2.

METHODSTo construct pEGFP-N1-IL2gpi plasmid coding a fusion gene of a DNA oligo encoding GPI-anchor signal sequence attaching to human IL-2 cDNA. Then T24 cell lines stably expressing GPI-IL-2 proteins (T24/GPI-IL-2) were established. Ex/GPI-IL-2 were isolated and purified by ultrafiltration and sucrose gradient centrifugation, and the morphology and molecule markers were analyzed. The mixed lymphocyte reaction study and cytotoxic study were performed to determine the proliferative effect of T lymphocytes and the cytotoxicity induced by Ex/GPI-IL-2.

RESULTSThe pEGFP-N1-IL2gpi plasmid was successfully constructed, and cell lines stably expressing GPI-IL-2 fusion proteins were established. Ex/GPI-IL-2 were small vesicular and saucer-shaped in diameter of 30-90 nm, containing heat shock protein 70, intercellular adhesion molecule-1, MAGE-1 and GPI-IL-2. Ex/GPI-IL-2-pulsed could dendritic cells induce proliferation of T cells and cytotoxic immune response more efficiently (P<0.05).

CONCLUSIONSGPI-IL-2 gene-modified tumor cells can make the exosomes containing GPI-IL-2 with an increased anti-tumor effect. Our study provides a feasible approach for exosome-based tumor immunotherapy of bladder transitional cell tumors.

Cancer Vaccines ; immunology ; Carcinoma, Transitional Cell ; immunology ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Exosomes ; genetics ; metabolism ; Glycosylphosphatidylinositols ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-2 ; metabolism ; Melanoma-Specific Antigens ; metabolism ; Plasmids ; Recombinant Fusion Proteins ; metabolism ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Urinary Bladder Neoplasms ; immunology ; metabolism ; pathology