Detection of TMPRSS2-ETS Fusions by a Multiprobe Fluorescence in Situ Hybridization Assay for the Early Diagnosis of Prostate Cancer Qi-Peng Sun, Liao-Yuan.

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Detection of TMPRSS2-ETS Fusions by a Multiprobe Fluorescence in Situ Hybridization Assay for the Early Diagnosis of Prostate Cancer Qi-Peng Sun, Liao-Yuan Li, Zhong Chen, Jun Pang, Wei-Jiao Yang, Xiang-Fu Zhou, Jian-Guang Qiu, Zu-Lan Su, Dan He, Xin Gao The Journal of Molecular Diagnostics Volume 12, Issue 5, Pages 718-724 (September 2010) DOI: 10.2353/jmoldx.2010.100002 Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 A schematic illustration of the design and labeling models for the ERG break-apart rearrangement probe (A), the TMPRSS2-ETV1 dual-color dual-fusion probe (B), and the TMPRSS2-ETV4 dual-color dual-fusion probe (C). The Journal of Molecular Diagnostics 2010 12, 718-724DOI: (10.2353/jmoldx.2010.100002) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 H&E stains and corresponding FISH images of the TMPRSS2-ETS fusion assay. A: Prostatic tissue with prostate cancer glands (Gleason score 5 + 3). B: FISH ERG probe image of the green boxed area in (A). The double-framed red inset is a magnification of the red boxed area showing three representative nuclei of a prostate cancer gland. Inset, Two nuclei with one yellow and one green signal, showing an ERG rearrangement involving partial deletion of the gene; a nucleus with one yellow, one green, and one red signal, showing an ERG rearrangement through translocation. C: Prostatic tissue with prostate cancer glands (Gleason score 2 + 2). D: FISH TMPRSS2-ETV1 probes image of the green boxed area in C. The double-framed red inset is a magnification of the red boxed area showing a representative nucleus with two yellow signals, indicating the TMPRSS2-ETV1 fusion. E: Prostatic tissue with prostate cancer glands (Gleason score 5 + 4). F: FISH TMPRSS2-ETV4 probes image of the green boxed area in E. The double-framed red inset is a magnification of the red boxed area showing a representative nucleus with two yellow signals, indicating the TMPRSS2-ETV4 fusion. G: Benign prostatic hyperplasia tissue. H: FISH ERG probe images of the green boxed areas in G. Each nucleus of the benign gland shows two yellow signals indicating absence of ERG related rearrangements. Original magnification of H&E images, ×20 objective. Original magnification of FISH images, oil objective (×100). The Journal of Molecular Diagnostics 2010 12, 718-724DOI: (10.2353/jmoldx.2010.100002) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 H&E stains, immunostaining, and corresponding FISH images of the TMPRSS2-ETS fusion assay in prostatic biopsy tissue. A: A prostatic gland with focal basal membrane disruption. B: FISH ERG probe images of the green boxed areas in A, demonstrating the presence of ERG-related rearrangements through partial deletion or translocation of the gene. C: Immunostaining of a prostatic gland in A by primary antibodies against 34βE12, p63, and AMACR, showing focal basal membrane disruption. D: An abnormal gland with focal basal membrane disruption surrounding benign prostatic glands. E: FISH TMPRSS2-ETV1 probes image of the green boxed area in D, indicating the presence of the TMPRSS2-ETV1 fusion. F: Immunostaining of a prostatic gland in D by primary antibodies against 34βE12, p63, and AMACR, showing focal basal membrane disruption. G: An abnormal gland with focal membrane disruption surrounding a benign prostatic gland. H: FISH TMPRSS2-ETV4 probes image of the green boxed area in E, indicating the presence of the TMPRSS2-ETV4 fusion. I: Immunostaining of a prostatic gland in G by primary antibodies against 34βE12, p63, and AMACR, showing focal basal membrane disruption. Original magnification of H&E images, ×20 objective. Original magnification of FISH images, oil objective (×100). The Journal of Molecular Diagnostics 2010 12, 718-724DOI: (10.2353/jmoldx.2010.100002) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions