Article Figures & Data
Figures
- Fig. 1.
Examples of HER-2 IHC staining of formalin-fixed, paraffin-embedded AD-CaP demonstrate visual scores of (A) 0, (B) 1+, (C) 2+, and (D) 3+ (reduced from magnification ×400).
- Fig. 2.
Quantitative RT-PCR. A, three-fold dilutions of HER-2 sRNA standard demonstrate dynamic range from 1,000 to 926,000 copies. B, linear regression demonstrates excellent linear fit (r = 0.99) for standard curve generated by 3-fold dilutions of HER-2 sRNA (•) standard. Also shown is distribution of tumor sample measurements (\#9676;) well within the range of the assay.
- Fig. 3.
Ratios of HER-2 copy number for AD-CaP versus patient matched benign prostate. No ratio is above 2 in contrast to experience with breast carcinomas, where HER-2 overexpression by IHC correlates with mRNA tumor/normal ratios of at least 4. Note that half of the matched prostate sets had ratios <1.
- Fig. 4.
FISH with CEP 17 (A, C, and E) and HER-2 probe (B, D, and F). For both probes, the negative controls are A and B, positive controls (E and F), and AI-CaP (C and D). The CaP specimen (HER-2/CEP 17 ratio = 1.40) demonstrated an intermediate ratio compared with an unamplified HER-2 (HER-2/CEP 17 ratio = 1.10) and weakly amplified HER-2 (HER-2/CEP 17 ratio = 1.78) controls.
Tables
- Table 1
Visual scoring of HER-2 immunostaining in AD-CaP versus AI-CaP
No. of patients 0 1+ 2+ 3+ AD-CaP 50 24 17 8 1 AI-CaP 25 24 1 0 0 - Table 2
Visual scoring of HER-2/neu immunostaining in AD-CaP classified by Gleason grade, pathologic stage, and race
No. of patients 0 1+ 2+ 3+ Gleason sum 5–6 17 9 4 4 0 7 24 9 12 3 0 8–10 9 6 1 1 1 Pathologic stage pT2 24 10 8 5 1 pT3 22 11 8 3 0 Node positive 4 3 1 0 0 Race Caucasian American 25 13 9 3 0 African American 25 11 8 5 1 - Table 3
HER-2 mRNA copies in benign, AD-CaP, and AI-CaP specimens
Tissue Type n Range Mean ± SD Benign 15 14,062 –30,574 23,957 ± 4,936 AD-CaPa 19 13,502 –39,856 22,080 ± 7,971 AI-CaPb 14 4,505 –22,073 15,496 ± 5,707 -
a P < 0.01, benign versus AD-CaP.
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b P < 0.05, AD-CaP versus AI-CaP.
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- Table 4
HER-2: CEP 17 ratios in AD-CaP and AI-CaP specimens from Caucasian (CA) and African (AA) Americans
Specimen HER-2: CEP 17 ratios AD-CaP/CA 1.07, 1.17, 1.31, 1.31, 1.48, 1.61, 1.63, 1.73, 1.78, 1.81 AD-CaP/AA 1.16, 1.19, 1.23, 1.25, 1.44, 1.44, 1.47, 1.51, 1.53, 1.56 AI-CaP/CA 1.02, 1.09, 1.26, 1.32, 1.37, 1.42, 1.49, 1.52, 1.62, 1.87 AI-CaP/AA 1.16, 1.16, 1.17, 1.17, 1.32, 1.34, 1.40, 1.48, 1.64 - Table 5
Review of HER-2 expression studies
Author Type Source Preservation Methods Antibody Overexpression definition % Overexpression Amplification definition Amplification Oxley 2001 (42) AD-CaP 108 AI-CaP 9 RPa Formalin-paraffin FISH NA NA NA 5 or more signals in >20% of nuclei 1.75% Liu 2001 (41) AD-CaP 30 RP Formalin-paraffin IHC FISH Dako Hercept CB11 (1:50) Novocastra 0–3% depending on which antibody was used 3 or more signals/ nucleus 53% (>3 spots/nucleus) Shi 2001 (43) AD-CaP 61 AI-CaP 20 RP TURP Formalin-paraffin IHC Dako (1:600) (hercept polyclonal) >50% cells with staining stronger than normal cells 29% of AD-CaP 50% of treated CaP 85% of AI-CaP NA NA Skacel 2001 (44) AD-CaP 39 RP Formalin-paraffin FISH NA NA NA 3–5 signals/nucleus 26% Osman 2001 (45) AD-CaP 83 AI-CaP 20 RP Mets. Formalin-paraffin IHC FISH Dako 485 Weak to moderate membrane staining in >10% of cells (2+) 38% >4 signals in ≥40% of nuclei 3% (2 of 66) Reese 2001 (47) AI-CaP 39 TURP Biopsy Formalin-paraffin IHC FISH TAB250 1:200 monoclonal 1+ weak 2+ moderate 3+ intense 10% (4 cases with 2+ or 3+ staining) HER-2/centromere 17 ratio ≥1.5 1 specimen Signoretti 2000 (46) AD-CaP 101 AI-CaP 18 RP 69 Formalin-paraffin IHC In situ hybri. FISH Dako 485 (1:200) Immunostaining > adjacent basal cells 25% of AD-CaP 59% of AI-CaP >4 signals/nucleus 0% Haussler 1999 (54) Adenosis 48 BPH 20 PIN 30 AD-CaP 38 RP TURP Formalin-paraffin IHC Dako 9G6 (1:600) monoclonal None, weak, moderate, and strong 0% of CaP 60% of PIN NA NA Morote 1999 (55) AI-CaP 70 Biopsy Formalin-paraffin IHC CB-11 (1:40) monoclonal Positive >1% of cells stained 64% NA NA Mydlo 1998 (39) AD-CaP 14 AI-CaP 3 RP 8 TURP 9 Formalin-paraffin IHC (CAS-200) 9G6 (1:2000) monoclonal CAS reading >0.06 pg/cell 3 of 17 revealed >20% of cells stained NA NA Ross 1997 (38) AD-CaP 113 RP Formalin-paraffin IHC (Dako) FISH Dako 485 (1:40) Average combined membranous and cytoplasmic stain >3 29% (intense cytoplasmic or diffuse membranous) 5 or more signals in >20% of nuclei 41% Kuhn 1993 (33) AD-CaP 53 RP Formalin-paraffin IHC pAb-1 (1:20) polyclonal Definite positive membranous staining 34% NA NA Mellon 1992 (37) AD-CaP 29 BPH 34 TURP Frozen IHC NCL-CB11 (1:40) (Novocastra) Grade 2 focal staining (1 = weak to moderate; 2 = strong) 21% of CaP NA NA Zhau 1992 (34) AD-CaP 16 BPH 6 NS Frozen IHC Western Ab-3 (5 ug/ml) 80% by IHC 68% by Western Southern blots 0% (0 of 10) Gu 1996 (32) AD-CaP 39 BPH 10 RP Formalin-paraffin IHC Ab-3 (1 ug/ml) 1, 10–20% cells 2, 20–50% cells 3, 50% cells 100% of CaP were 1–3 20% of BPH NA NA Visakorpi 1992 (57) AD-CaP 147 BPH 17 Biopsy 101 TURP 46 Formalin-paraffin IHC Mab-1 NS 0% NA NA Veltri 1994 (40) AD-CaP 124 RP Formalin-paraffin IHC Ab-3 Focal, diffuse, or none 78% NA NA Sandavisan 1993 (31) AD-CaP 25 BPH 15 RP TURP Frozen IHC TA-1 (0.5 ug/ml) monoclonal 1+ <5% 2+ <90% 3+ >90% 36% had 3+ NA NA -
a ABC, avidin-biotin peroxidase; IAP, immunoalkaline phosphatase; PIN, prostatic epithelial neoplasia; TURP, Transurethral resection of prostate; RP, radical prostatectomy; Mets, metastases; NA, not applicable; NS, not specified.
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Volume 9, Issue 3
