Table 1.
Summary of Immunohistochemical Analysis of Ep-ICD in Normal and Epithelial Cancers.
Figure 1.
Immunohistochemical analysis of Ep-ICD expression in epithelial cancers and normal tissues.
The representative photomicrographs depict Ep-ICD immunostaining in normal and cancer tissues. Panel A shows predominant membrane localization of Ep-ICD and no nuclear staining in the normal breast tissue (I), while the cancer tissue shows nuclear and cytoplasmic Ep-ICD accumulation (II). Panel B shows low level of membrane Ep-ICD in the epithelial cells and the basal cells show some nuclear staining in the normal prostate tissue (Ia) and in benign prostate hyperplasia (B, Ib), while the cancer tissue shows intense cytoplasmic and nuclear staining (II). Panel C shows no detectable Ep-ICD staining in the normal esophageal tissue (I), while the ESCC shows intense nuclear and cytoplasmic immunostaining (II). Panel D depicts head and neck normal mucosa showing faint membrane Ep-ICD (I), while the HNSCC shows intense nuclear and cytoplasmic immunostaining (II). Original magnification ×400.
Figure 2.
Immunohistochemical analysis of Ep-ICD expression in epithelial cancers.
Increased nuclear and cytoplasmic immunoreactivity and reduced or absence of membrane staining of Ep-ICD was observed in cancers of the bladder (Panel A), lung (Panel B), liver (Panel C), ovary (Panel D), colon (Panel E), and pancreas (Panel F). Original magnification ×400.
Figure 3.
Immunohistochemical analysis of Ep-ICD expression in prostate and colon cancers.
Membrane, cytoplasmic, and nuclear Ep-ICD expression in prostate cancer (A) and colon cancer (B). Some areas of the tissue section show predominant membrane and weak cytoplasmic but no nuclear localization of Ep-ICD (A, B - left square), while the other areas of the same tissue section show nuclear and cytoplasmic accumulation of Ep-ICD with absence of membranous Ep-ICD expression (A, B - right square). Original magnification ×400.
Figure 4.
Ep-ICD immunohistochemical analysis in epithelial cancers control tissues.
The negative and positive control photomicrographs are shown. Ep-ICD negative controls for HNSCC (A), prostate cancer (B), colon cancer (C), breast cancer (D), ESCC (E), and normal esophagus (F); panels G and H are positive controls for Ep-ICD staining. Original magnification ×400.
Figure 5.
Scatter plot analysis of Ep-ICD membrane, cytoplasmic, and nuclear expression.
Scatter plots showing distribution of total immunostaining scores determined by IHC of tissue sections of cancers of the breast (n = 38), prostate (n = 49), lung (n = 59), ovary (n = 10), colon (n = 59), bladder (n = 10), liver (n = 9), positively-staining HNSCCs (n = 39) and ESCCs (n = 19), and normal breast (n = 25), prostate (n = 9) and BPH (n = 21), normal esophageal (n = 20), and head and neck (n = 20) tissues. The vertical axis gives the total IHC score as described in the Methods. A cutoff of ≥4 was used to determine positivity. N, normal; Ca, cancer. A. Increased nuclear accumulation of Ep-ICD was observed in most of the epithelial cancers analyzed. B. Increased cytoplasmic accumulation of Ep-ICD was observed in almost all epithelial cancers analyzed. C. Membrane localization of Ep-ICD varied in the different cancer and normal tissue types examined.
Figure 6.
Immunohistochemical analysis of EpEx expression in epithelial cancers.
The photomicrographs depict MOC31 stained membrane EpEx in epithelial cancers. The panel I shows low level of membrane EpEx expression in normal breast (A) and prostate (B, Ia), BPH (B, Ib), and normal esophagus (C) and head and neck (D) tissues. The corresponding cancer tissues depicting increased level of EpEx in the membrane are shown in panel II (A–D). In contrast many of the cancer tissues of each cancer type showed absence of membrane EpEx (panel III, A–D). Original magnification ×400.
Figure 7.
Immunohistochemical analysis of EpEx expression in epithelial cancers.
Membrane EpEx expression was observed in all the epithelial cancers. Panel I shows intense membrane EpEx in colon cancer (A), liver (B), bladder (C), lung (D), ovarian (E) and pancreatic (F) cancer. The panels II A–F show reduced or absence of membrane EpEx in a subset of each of these epithelial cancers. Original magnification ×400.
Figure 8.
Fluorescence immunostaining in CX-1 cells with anti-EpEx (MOC31) and anti-Ep-ICD antibodies.
Secondary antibodies are FITC-anti-mouse (green) and TRITC-anti-rabbit (red). A) EpEx; B) Ep-ICD; C) DAPI; D) EpEx and DAPI (A&C merged); E) Ep-ICD and DAPI (B & C merged); F) EpEx and Ep-ICD (A & B merged); G) EpEx, Ep-ICD, and DAPI (A, B, C merged); I) Measurement of the density of three colors across the line in the cells in H.
Figure 9.
ROC curves of nuclear and cytoplasmic Ep-ICD in prostate cancer, breast cancer, HNSCC, and ESCC.
ROC curves describing the relationship between sensitivities and 1-specificities of nuclear and cytoplasmic Ep-ICD expression in these epithelial cancers. The vertical axis of each curve indicates sensitivity and the horizontal axis indicates the 1-specificity. The sensitivity, specificity, and AUC values for the cancers are summarized in Table 2.
Table 2.
Biomarker Analysis of Nuclear and Cytoplasmic Ep-ICD Expression in Epithelial Cancers.