Human Prostate Epithelial Cells

Purpose Bladder outlet obstruction (BOO) leads to detrusor overactivity (DO) and structural remodeling of the bladder. However, the molecular mechanisms underlying this ... More

Purpose Bladder outlet obstruction (BOO) leads to detrusor overactivity (DO) and structural remodeling of the bladder. However, the molecular mechanisms underlying this process remain incompletely understood. Materials and Methods In a rat model of partial BOO, we assessed bladder function via cystometry and evaluated histological changes using hematoxylin and eosin staining. The expression and localization of aquaporin (AQP)-2 and caveolin (CAV)-1, -2, and -3 were analyzed by Western blotting and immunofluorescence in bladder tissues. Additionally, immunostaining was performed in cultured human bladder smooth muscle cells to assess protein co-localization. Results BOO rats exhibited elevated detrusor contraction pressure and shortened intercontraction intervals. Histologically, hypertrophied smooth muscle bundles and increased extracellular matrix were observed. Western blot analysis revealed significant upregulation of AQP2 and CAV1–3 in BOO bladders. Tissue immunofluorescence demonstrated increased expression and redistribution of these proteins in the detrusor muscle. Cultured cell analysis confirmed subcellular co-localization of AQP2 with CAV1–3. Conclusions DO induced by BOO is associated with upregulated expression of AQP2 and CAV1–3 in the urinary bladder. The coexpression observed in bladder cells suggests potential molecular interactions between AQP2 and CAVs that may contribute to the pathophysiology of BOO-related bladder dysfunction. Keywords: Aquaporin-2; Bladder outlet obstruction; Caveolin; Urinary bladder, overactive Less

This study reports on probing the utility of in situ chromatin texture features such as nuclear DNA methylation and chromatin condensation patterns — visualized by fluo... More

This study reports on probing the utility of in situ chromatin texture features such as nuclear DNA methylation and chromatin condensation patterns — visualized by fluorescent staining and evaluated by dedicated three-dimensional (3D) quantitative and high-throughput cell-by-cell image analysis — in assessing the proliferative capacity, i.e. growth behavior of cells: to provide a more dynamic picture of a cell population with potential implications in basic science, cancer diagnostics/prognostics and therapeutic drug development. Two types of primary cells and four different cancer cell lines were propagated and subjected to cell-counting, flow cytometry, confocal imaging, and 3D image analysis at various points in culture. Additionally a subset of primary and cancer cells was accelerated into senescence by oxidative stress. DNA methylation and chromatin condensation levels decreased with declining doubling times when primary cells aged in culture with the lowest levels reached at the stage of proliferative senescence. In comparison, immortal cancer cells with constant but higher doubling times mostly displayed lower and constant levels of the two in situ-derived features. However, stress-induced senescent primary and cancer cells showed similar levels of these features compared with primary cells that had reached natural growth arrest. With regards to global DNA methylation and chromatin condensation levels, aggressively growing cancer cells seem to take an intermediate level between normally proliferating and senescent cells. Thus, normal cells apparently reach cancer-cell equivalent stages of the two parameters at some point in aging, which might challenge phenotypic distinction between these two types of cells. Companion high-resolution molecular profiling could provide information on possible underlying differences that would explain benign versus malign cell growth behaviors. Keywords: DNA methylation, chromatin condensation, cell proliferation, aging, senescence, cancer, 3D imaging, cell-by-cell analysis Less