43 research outputs found

    Morphologic changes in the derivative clones during colony formation.

    No full text
    <p>The morphology of a derivative colony was followed during the cloning process, growing from a single well of a 96-well plate to a single well of 24-well plate, to a single well of a 6-well plate, and to a 10 cm dish. For each view, a phase contrast image (top) and red fluorescence image (bottom) are shown. All the images are shown at 100× magnification.</p

    Confirming cancer-stromal cell fusion.

    No full text
    <p>Representative cancer-stromal cell fusion events from the co-culture of RL-1 cells with hMSC-GFP cells are shown. <b>A</b>, A single fusion event at day 7 is shown in bright field, green fluorescence, and red fluorescence. The green and red fluorescence images are merged (merged fluorescence) to show the two nuclei of different fluorescence. <b>B</b>, Merged fluorescence images for 4 additional fusion events are shown, with events 1 and 2 recorded at day 7, and 3 and 4 at day 14. Arrows are used to indicate nuclei. All the images are shown at 200× magnification.</p

    Genotypic and phenotypic changes in the derivative clones from cancer-stromal fusion.

    No full text
    <p>Genotypic and phenotypic parameters of the first 9 clones of the RL-1 and HPS-15 fusion hybrids were compared to those of the first 12 clones from control cloning. Compared to RL-1 clones, all the derivative clones lost Y chromosomes (<b>A</b> versus <b>B</b>). Detected by Western blotting, some of the derivative clones showed persistent AR expression even under androgen-deprivaton (<b>C</b> versus <b>D</b>). In these studies, cells were cultured for 48 hours in regular culture medium (C), androgen deprivation medium (−), and androgen deprivation medium containing 5 nM R1881 (+). The derivative clones were detected to express increased levels of PSA, even during androgen-deprivation (<b>E</b> versus <b>F</b>). When growth rate was assayed by MTT conversion, clones derived from cancer-stromal fusion displayed accelerated growth in androgen-independent fashion (<b>G</b> versus <b>H</b>). Data represent the mean of triplicate assays. For all the data points, standard deviation was less than 5% of the mean and is not shown.</p

    Tracking the fate of cancer-stromal hybrids.

    No full text
    <p>Representative morphologies of the hybrid cells during colony formation are shown. <b>A</b>, Two weeks into the culture, most of the hybrids contained two nuclei of similar fluorescence. No cell division was observed. <b>B</b>, Four weeks into the co-culture, hybrid cells adopted atypical morphology with multiple nuclei. No cell division was observed. <b>C</b>, Six weeks into the culture, the remaining hybrid cells became thin or narrow, with multiple nuclei in segments of the cell. <b>D</b>, Eight weeks into the culture, cell division became prevalent. The cell division was abnormal because it produced daughter cells in varied shapes and with reduced viability. For each view, a phase contrast image (top) and red fluorescence image (bottom) are shown. When necessary, arrows are used to indicate nuclei. All the images are shown at 200× magnification.</p

    Reduced colony formation in cancer-stromal fusion hybrids.

    No full text
    a<p>Wells containing a single cell 24 hours after the plating were enumerated.</p>b<p>Colonies from the wells containing a single cell were enumerated.</p>c<p>Data were from one colony formation assay.</p>d<p>Data were combined results from 4 repeated colony formation assays.</p

    Time-dependence of cancer-stromal cell fusion.

    No full text
    <p>Co-cultures of RL-1 and HPS-15 cells were observed weekly for frequency of cell fusion. For each view, a phase contrast image (top) and red fluorescence image (bottom) are shown. Arrows are used to indicate cancer-stromal cell fusion events. All the images are shown at 40× magnification.</p

    Characteristics of the spontaneous cancer-stromal cell fusion.

    No full text
    <p>RL-1 cells (<b>A</b>) and HPS-15 cells (<b>B</b>) are shown in separate culture. After 7 days of co-culture, spontaneous fusion could be seen (<b>C</b>). At higher magnification, the fused cell contained two nuclei, one fluorescently red and the other fluorescently pale (<b>D</b>). Cancer-stromal fusion was frequently seen in areas where RL-1 and HPS-15 formed close contact (<b>E</b>). In some cases, cells in the middle of a fusion could be seen (<b>F</b>). The two nuclei could be seen close to each other (<b>G</b>) or separated (<b>H</b>). For each view, a phase contrast image (top) and red fluorescence image (bottom) are shown. Arrows are used to indicate nuclei.</p

    Multiplexed Quantum Dot Labeling of Activated c-Met Signaling in Castration-Resistant Human Prostate Cancer

    No full text
    The potential application of multiplexed quantum dot labeling (MQDL) for cancer detection and prognosis and monitoring therapeutic responses has attracted the interests of bioengineers, pathologists and cancer biologists. Many published studies claim that MQDL is effective for cancer biomarker detection and useful in cancer diagnosis and prognosis, these studies have not been standardized against quantitative biochemical and molecular determinations. In the present study, we used a molecularly characterized human prostate cancer cell model exhibiting activated c-Met signaling with epithelial to mesenchymal transition (EMT) and lethal metastatic progression to bone and soft tissues as the gold standard, and compared the c-Met cell signaling network in this model, in clinical human prostate cancer tissue specimens and in a castration-resistant human prostate cancer xenograft model. We observed c-Met signaling network activation, manifested by increased phosphorylated c-Met in all three. The downstream survival signaling network was mediated by NF-kB and Mcl-1 and EMT was driven by receptor activator of NF-kB ligand (RANKL), at the single cell level in clinical prostate cancer specimens and the xenograft model. Results were confirmed by real-time RT-PCR and western blots in a human prostate cancer cell model. MQDL is a powerful tool for assessing biomarker expression and it offers molecular insights into cancer progression at both the cell and tissue level with high degree of sensitivity
    corecore