lucifer-yellow and Prostatic-Neoplasms

This paper examines and compares necrosis in human ovarian tissue after conventional slow freezing or vitrification and ensuing xenotranplantation. Slow cryoconserved or vitrified ovarian tissue samples and fresh controls from nine patients were subcutaneously transplanted into SCID mice. The tissue samples were explanted after 6 weeks and the necrotic areas were examined by staining with Lucifer yellow SV. The size of the necrotic areas in parallel cultivated ovarian tissue samples was compared, as was necrosis in cultivated prostate tumour spheroids where the emergence of necrosis and its pathophysiological correlation have been described. Examinations showed no significant rise in the proportion of necrotic areas after slow cryoconservation/transplantation and in the controls (transplanted fresh tissue, not transplanted fresh tissue, long-term culture). The proportion of necrotic areas in the tumour spheroids was significantly higher than in the ovarian tissue. Vitrification could, after these results, be presented as an alternative to conventional slow cryoconservation.

Topics: Adult; Animals; Cell Line, Tumor; Cryopreservation; Female; Fluorescent Dyes; Freezing; Humans; Isoquinolines; Male; Mice; Mice, SCID; Necrosis; Organ Transplantation; Organic Chemicals; Ovary; Prostatic Neoplasms; Time Factors; Transplantation, Heterologous

Gap junctional communication (GJC) has been implicated in the control of cell proliferation. Numerous cancer cells show a decrease or loss of GJC compared to their normal counterparts. Lack of adequate information on the status of gap junctions during prostate neoplasia prompted us to examine this form of cancer, which comprises about 14% of male cancer deaths in America.. Cultured normal human prostate epithelial cells and several different human prostate tumor lines were used in this study. GJC was assayed by dye transfer, whereas Western blot and immunofluorescence methods were used to examine connexin43 (Cx43) levels and the presence of gap junctions, respectively.. Normal human prostate cultures exhibited extensive cell-communication which was completely absent in all the examined tumor cells. This disrupted communication was associated with a decreased expression and an impaired posttranslational modification of Cx43 in these cells. Abundant immunostaining of gap junctional channels by a Cx43-antibody was observed in normal prostate cells but not in tumor cells.. Our data provide further support for the hypothesis that loss of junctional communication is a critical step in progression to human prostate neoplasia.

Topics: Blotting, Western; Cell Communication; Connexin 43; Down-Regulation; Fluorescent Antibody Technique; Fluorescent Dyes; Gap Junctions; Gene Expression Regulation, Neoplastic; Humans; Isoquinolines; Male; Prostatic Neoplasms; Protein Processing, Post-Translational; Tumor Cells, Cultured

To investigate gap-junctional intercellular communication (GJIC) in LNCaP and DU145 human prostate cancer cells.. Normal rat liver F344 (WB1) cells were used as positive controls. Functional GJIC was inspected using either the scrape-loading/dye transfer (SL/DT) method or fluorescence recovery after photobleaching (FRAP) analysis. In the former, GJIC activity was expressed as a measure of the extent of diffusion of Lucifer Yellow after cell monolayers were scraped using a surgical blade and exposed to dye for a few minutes at room temperature. In the latter, cells were incubated for 15 minutes at 37 degrees C with 5,6-carboxyfluorescein diacetate dye and the dye transfer visualized by photobleaching individual cells with a 488-nm laser and monitoring the recovery of fluorescence using a laser cytometer.. The preliminary results obtained indicate that neither LNCaP nor DU145 cells have functional GJIC, while, as expected, WB1 cells show unimpaired GJIC activity. Equivalent results were consistently obtained using either SL/DT or the FRAP approach. However, using FRAP analysis, DU145 cells only showed weak recovery of fluorescence after a total observation interval of 15 minutes.. The present data, though preliminary, suggest that disruption of GJIC may play a role in development of malignancy in the human prostate.

Topics: Animals; Cell Communication; Fluoresceins; Fluorescent Dyes; Gap Junctions; Humans; Isoquinolines; Male; Microscopy, Confocal; Microscopy, Fluorescence; Prostatic Neoplasms; Rats; Tumor Cells, Cultured