EpCAM expression of human malignant cells.

<p>(A) The mRNA of the malignant cell lines 5061, 5072 (pancreatic cancer), LNCAP, PC3 (prostate cancer), FemX-1, MEWO (melanoma), T47D, MCF7 (breast cancer), HT29 (colon cancer) and OH-1 (small cell lung cancer) were relatively quantified by qPCR, using GAPDH for normalization. 5072, LNCAP, PC3, T47D, MCF7, Caco2 and HT29 showed high expression levels of EpCAM mRNA. (B) EpCAM could be detected by Western blot analysis of HT29 cell lysate with a specific binding to antibody MOC31. Beta-actin was used as loading control. (C) EpCAM could positively be detected by flow cytometry analysis with MOC31 on all cancer cell lines, except of FemX-1 and MEWO. Isotype controls are shown as dotted lines.</p

MRI of HT29 xenograft.

<p>HT29 colon carcinoma (a) grown at the subcutaneous injection site above muscles (b) appear hyper-intense in two-dimensional turbo spin-echo (TSE) sequence (MR images in axial orientation).</p

IFP measurements of HT29 xenograft tumours.

<p>(A) Typical pressure recording of a HT29 tumour. After adjusting the pressure at 0 mbar and insertion of the needle (arrow a) in the tumour the pressure increased and decreased rapidly and reaches a stable value of 16 mbar. To test the fluid communication between pressure sensor and needle the tubing is compressed with a screw clamp (arrow b) which results in a sharp increase of the pressure, then an exponential decrease, followed by another period of stabilization before decompressing the tubing (arrow c) which produces a rapid decrease and an exponential increase. (B) Measurement of 6 different HT29 tumours showed a mean IFP of 11.9 (+/−4) mbar.</p

EpCAM <i>in vivo</i> binding.

<p>[125I]-Labelled specific anti EpCAM MOC31 and non-specific [125I]-labelled IgG1 antibody were used for EpCAM <i>in vivo</i> binding in HT29 carcinoma bearing SCID mice. There is a significant difference between specific and control antibody by HT29 carcinoma, but except for spleen and blood not in other organs (two-way ANOVA, P<0.001, n = 3). Standard deviations are indicated by bars.</p

Additional file 1 of PSMA-ligand uptake can serve as a novel biomarker in primary prostate cancer to predict outcome after radical prostatectomy

Additional file 1. Supplementary files. Supplementary table 1. Distribution of pT and miT. Supplementary table 2. Distribution of pN and miN. Supplementary table 3. Univariate analysis for the association of 68Ga-PSMA-11 PET findings with surgical margin status. Supplementary Fig. 1. Flowchart of inclusion and exclusion steps. Supplementary Fig. 2. Longer biochemical recurrence-free survival was associated with (A) pT = 2, (B) pN=0, (C) Gleason Score < 8 and (D) negative surgical margin

Evans Blue-Albumin complex distribution in vibratome sections of HT29 carcinoma.

<p>Evans Blue-Albumin complex positive areas (blue) are recognizable at blood vessels in vital tumour tissue (b), to a higher degree at well perfused areas at the margin of the tumour (a) and in the transition between vital tumour tissue and necrosis (c).</p

EpCAM <i>in vivo</i> detection of HT29 tumour.

<p>MOC31-antibody binds to HT29 tumour cells <i>in vivo</i> after i.v. injection of 10 µg MOC31mAb in tumour bearing mice, as visualized with subsequent immunostaining against MOC31-antibody (red = MOC31 positive cells) in cryostat sections of the primary tumour. Controls using 10 µg IgG1 mAb confirmed the high specificity of MOC31 binding to tumour cells <i>in vivo</i>. The specific antibody MOC31 binding to the tumour cells was restricted to areas of the tumour, which were well supplied with blood vessels. Corresponding liver sections showed the intake of both antibodies by cells of the RES system.</p

Vasculature of HT29 xenografted tumours visualized by DiI labelling.

<p>(A) Unequal distribution of the blood vasculature (red) of a whole HT29 tumour vibratome slice (stitched of ×10 objective lens images). (B) Magnification of the boxed area in A, showing immature blood vessels (projection of a z series with z step size of 2.85 µm over 215 µm with ×10 objective lens). (C) Three-dimensional view of a section of B (z series with z step size of 1.4 µm over 140 µm with ×40 objective lens) showing the irregular structure of blood vessels. All tissues were viewed by confocal fluorescence microscopy (Nikon A1R confocal microscope with plan apo ×10/0.45 numerical aperture (NA) or plan fluo oil ×40/1.3 NA objective lens and with the laser excitation wavelength of 561 nm and the emission of 595). Scale bars: 2000 µm for A, 200 µm for B.</p

EpCAM protein expression pattern <i>in vivo</i> and <i>in vitro</i>.

<p>5061, LNCAP, PC3, T47D, HT29, and Caco2 cells showed <i>in vitro</i> and <i>in vivo</i> strong binding of MOC31 by all cells. In contrast, MCF7, OH-1, SW480 and particularly 5072 cells showed strong MOC31 <i>in vitro</i> binding, but little or no binding <i>in vivo</i>. MEWO and FemX-1 cells showed no MOC31 binding in vitro and in vivo. (red = MOC31 binding).</p

Summary of <i>in vitro</i> and <i>in vivo</i> EpCAM expression of all cell lines by different methods.

<p>Note that the highest EpCAM expression both <i>in vitro</i> and <i>in vivo</i> was observed in the cell lines 5061, LNCAP, PC3, HT29, and Caco2. The intensity (plus signs) and extent (percent) of the positive areas of 5 histological sections were determined by visual inspection of 3 independent observers.</p