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TEAD1 and c-Cbl are novel prostate basal cell markers that correlate with poor clinical outcome in prostate cancer.

Knight JF, Shepherd CJ, Rizzo S, Brewer D, Jhavar S, Dodson AR, Cooper CS, Eeles R, Falconer A, Kovacs G, Garrett MD, Norman AR, Shipley J, Hudson DL - Br. J. Cancer (2008)

Bottom Line: RNA from basal and luminal cells isolated from benign tissue by immunoguided laser-capture microdissection was subjected to expression profiling.We identified 112 and 267 genes defining basal and luminal populations, respectively.Analyses of prostate cancer tissue microarray staining established that increased protein levels of either marker were associated with decreased patient survival independent of other clinicopathological metrics.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Carcinogenesis, The Bob Champion Prostate Stem Cell Team, The Institute of Cancer Research, Sutton, Surrey SM2 5NG, UK.

ABSTRACT
Prostate cancer is the most frequently diagnosed male cancer, and its clinical outcome is difficult to predict. The disease may involve the inappropriate expression of genes that normally control the proliferation of epithelial cells in the basal layer and their differentiation into luminal cells. Our aim was to identify novel basal cell markers and assess their prognostic and functional significance in prostate cancer. RNA from basal and luminal cells isolated from benign tissue by immunoguided laser-capture microdissection was subjected to expression profiling. We identified 112 and 267 genes defining basal and luminal populations, respectively. The transcription factor TEAD1 and the ubiquitin ligase c-Cbl were identified as novel basal cell markers. Knockdown of either marker using siRNA in prostate cell lines led to decreased cell growth in PC3 and disrupted acinar formation in a 3D culture system of RWPE1. Analyses of prostate cancer tissue microarray staining established that increased protein levels of either marker were associated with decreased patient survival independent of other clinicopathological metrics. These data are consistent with basal features impacting on the development and clinical course of prostate cancers.

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Immunofluorescent and immunohistochemical labelling of benign and cancerous prostate tissue for basal and luminal cell markers. (A) Benign tissue: nuclear TEAD1 labelling (green) was found in K14-positive basal cells (red). Note turquoise double-stained TEAD1-positive nuclei. Cytoplasmic staining for c-Cbl (green) was strong in basal cells and weak in luminal cells as shown by co-labelling with luminal marker K8 (red). SNAP25 (green) staining was luminal in a speckled vesicle-like pattern. Integrin αV (green) was restricted to basal cells, either co-localised to K14 (red) or alone. In prostate tumours, the expression of TEAD1 and c-Cbl was strong despite the absence of a basal layer. Occasional areas of tumour tissue labelled intensely for SNAP25. Integrin αV expression in tumour tissue was extremely weak. Original magnification × 63 to × 100. (B) Scoring systems were derived for both TEAD1 and c-Cbl. TEAD1 scored first as focal (F) or diffuse (D) followed by a score for intensity (1=low to 3=high). Tumour cores lacking TEAD1 expression were scored as negative (neg). Scoring for c-Cbl was based on intensity alone (C1=low to C 3=high). Original magnification × 40.
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fig2: Immunofluorescent and immunohistochemical labelling of benign and cancerous prostate tissue for basal and luminal cell markers. (A) Benign tissue: nuclear TEAD1 labelling (green) was found in K14-positive basal cells (red). Note turquoise double-stained TEAD1-positive nuclei. Cytoplasmic staining for c-Cbl (green) was strong in basal cells and weak in luminal cells as shown by co-labelling with luminal marker K8 (red). SNAP25 (green) staining was luminal in a speckled vesicle-like pattern. Integrin αV (green) was restricted to basal cells, either co-localised to K14 (red) or alone. In prostate tumours, the expression of TEAD1 and c-Cbl was strong despite the absence of a basal layer. Occasional areas of tumour tissue labelled intensely for SNAP25. Integrin αV expression in tumour tissue was extremely weak. Original magnification × 63 to × 100. (B) Scoring systems were derived for both TEAD1 and c-Cbl. TEAD1 scored first as focal (F) or diffuse (D) followed by a score for intensity (1=low to 3=high). Tumour cores lacking TEAD1 expression were scored as negative (neg). Scoring for c-Cbl was based on intensity alone (C1=low to C 3=high). Original magnification × 40.

Mentions: Commercially available antibodies were used to confirm the protein localisation of a selection of novel markers in benign and malignant prostate tissue by immunofluorescence. The selection included genes identified as basal (ITGAV, TEAD1, c-Cbl and IL6) and luminal (SPRY1 and SNAP25). Double labelling with either K14 or K8 was used to confirm basal or luminal localisation of staining within benign prostate tissue. In agreement with the expression profiling data, TEAD1, c-Cbl and integrin αV were expressed in the basal layer and SNAP25 in the luminal layer (Figure 2A). We were unable to detect staining for IL6 or SPRY1. TEAD1 staining was nuclear in all cells within the basal layer, whereas c-Cbl was strongly cytoplasmic in the basal layer, with weak luminal staining. Integrin αV was found in subsets of cells within the basal layer, and clusters of integrin αV-positive cells were either K14 positive or negative. The SNAP25 antibody detected luminal cells, with a vesicular staining pattern. The same antibodies were also used on a small selection of prostate tumour samples. Although expression of SNAP25 and integrin αV could be detected in the tumours, integrin αV labelling was often weak, and results for SNAP25 were inconsistent between samples. As c-Cbl and TEAD1 staining was consistently high, these proteins were chosen for further study. c-Cbl was expressed in the cytoplasm of epithelial tumour cells, whereas TEAD1 was confined to the nuclei (Figure 2A).


TEAD1 and c-Cbl are novel prostate basal cell markers that correlate with poor clinical outcome in prostate cancer.

Knight JF, Shepherd CJ, Rizzo S, Brewer D, Jhavar S, Dodson AR, Cooper CS, Eeles R, Falconer A, Kovacs G, Garrett MD, Norman AR, Shipley J, Hudson DL - Br. J. Cancer (2008)

Immunofluorescent and immunohistochemical labelling of benign and cancerous prostate tissue for basal and luminal cell markers. (A) Benign tissue: nuclear TEAD1 labelling (green) was found in K14-positive basal cells (red). Note turquoise double-stained TEAD1-positive nuclei. Cytoplasmic staining for c-Cbl (green) was strong in basal cells and weak in luminal cells as shown by co-labelling with luminal marker K8 (red). SNAP25 (green) staining was luminal in a speckled vesicle-like pattern. Integrin αV (green) was restricted to basal cells, either co-localised to K14 (red) or alone. In prostate tumours, the expression of TEAD1 and c-Cbl was strong despite the absence of a basal layer. Occasional areas of tumour tissue labelled intensely for SNAP25. Integrin αV expression in tumour tissue was extremely weak. Original magnification × 63 to × 100. (B) Scoring systems were derived for both TEAD1 and c-Cbl. TEAD1 scored first as focal (F) or diffuse (D) followed by a score for intensity (1=low to 3=high). Tumour cores lacking TEAD1 expression were scored as negative (neg). Scoring for c-Cbl was based on intensity alone (C1=low to C 3=high). Original magnification × 40.
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Related In: Results  -  Collection

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fig2: Immunofluorescent and immunohistochemical labelling of benign and cancerous prostate tissue for basal and luminal cell markers. (A) Benign tissue: nuclear TEAD1 labelling (green) was found in K14-positive basal cells (red). Note turquoise double-stained TEAD1-positive nuclei. Cytoplasmic staining for c-Cbl (green) was strong in basal cells and weak in luminal cells as shown by co-labelling with luminal marker K8 (red). SNAP25 (green) staining was luminal in a speckled vesicle-like pattern. Integrin αV (green) was restricted to basal cells, either co-localised to K14 (red) or alone. In prostate tumours, the expression of TEAD1 and c-Cbl was strong despite the absence of a basal layer. Occasional areas of tumour tissue labelled intensely for SNAP25. Integrin αV expression in tumour tissue was extremely weak. Original magnification × 63 to × 100. (B) Scoring systems were derived for both TEAD1 and c-Cbl. TEAD1 scored first as focal (F) or diffuse (D) followed by a score for intensity (1=low to 3=high). Tumour cores lacking TEAD1 expression were scored as negative (neg). Scoring for c-Cbl was based on intensity alone (C1=low to C 3=high). Original magnification × 40.
Mentions: Commercially available antibodies were used to confirm the protein localisation of a selection of novel markers in benign and malignant prostate tissue by immunofluorescence. The selection included genes identified as basal (ITGAV, TEAD1, c-Cbl and IL6) and luminal (SPRY1 and SNAP25). Double labelling with either K14 or K8 was used to confirm basal or luminal localisation of staining within benign prostate tissue. In agreement with the expression profiling data, TEAD1, c-Cbl and integrin αV were expressed in the basal layer and SNAP25 in the luminal layer (Figure 2A). We were unable to detect staining for IL6 or SPRY1. TEAD1 staining was nuclear in all cells within the basal layer, whereas c-Cbl was strongly cytoplasmic in the basal layer, with weak luminal staining. Integrin αV was found in subsets of cells within the basal layer, and clusters of integrin αV-positive cells were either K14 positive or negative. The SNAP25 antibody detected luminal cells, with a vesicular staining pattern. The same antibodies were also used on a small selection of prostate tumour samples. Although expression of SNAP25 and integrin αV could be detected in the tumours, integrin αV labelling was often weak, and results for SNAP25 were inconsistent between samples. As c-Cbl and TEAD1 staining was consistently high, these proteins were chosen for further study. c-Cbl was expressed in the cytoplasm of epithelial tumour cells, whereas TEAD1 was confined to the nuclei (Figure 2A).

Bottom Line: RNA from basal and luminal cells isolated from benign tissue by immunoguided laser-capture microdissection was subjected to expression profiling.We identified 112 and 267 genes defining basal and luminal populations, respectively.Analyses of prostate cancer tissue microarray staining established that increased protein levels of either marker were associated with decreased patient survival independent of other clinicopathological metrics.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Carcinogenesis, The Bob Champion Prostate Stem Cell Team, The Institute of Cancer Research, Sutton, Surrey SM2 5NG, UK.

ABSTRACT
Prostate cancer is the most frequently diagnosed male cancer, and its clinical outcome is difficult to predict. The disease may involve the inappropriate expression of genes that normally control the proliferation of epithelial cells in the basal layer and their differentiation into luminal cells. Our aim was to identify novel basal cell markers and assess their prognostic and functional significance in prostate cancer. RNA from basal and luminal cells isolated from benign tissue by immunoguided laser-capture microdissection was subjected to expression profiling. We identified 112 and 267 genes defining basal and luminal populations, respectively. The transcription factor TEAD1 and the ubiquitin ligase c-Cbl were identified as novel basal cell markers. Knockdown of either marker using siRNA in prostate cell lines led to decreased cell growth in PC3 and disrupted acinar formation in a 3D culture system of RWPE1. Analyses of prostate cancer tissue microarray staining established that increased protein levels of either marker were associated with decreased patient survival independent of other clinicopathological metrics. These data are consistent with basal features impacting on the development and clinical course of prostate cancers.

Show MeSH
Related in: MedlinePlus