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Tumour-initiating stem-like cells in human prostate cancer exhibit increased NF-κB signalling.

Rajasekhar VK, Studer L, Gerald W, Socci ND, Scher HI - Nat Commun (2011)

Bottom Line: These TICs possess stem cell characteristics and multipotency as demonstrated by in vitro sphere-formation and in vivo tumour-initiation, respectively.The cells represent an undifferentiated subtype of basal cells and can be purified from prostate tumours based on coexpression of the human pluripotent stem cell marker TRA-1-60 with CD151 and CD166.These TICs exhibit increased nuclear factor-κB activity.

View Article: PubMed Central - PubMed

Affiliation: 1] Stem Cell Center and Developmental Biology Program, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA. [2] Sidney Kimmel Center for Prostate and Urologic Cancers, Department of Medicine, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA.

ABSTRACT
Androgen depletion is a key strategy for treating human prostate cancer, but the presence of hormone-independent cells escaping treatment remains a major therapeutic challenge. Here, we identify a minor subset of stem-like human prostate tumour-initiating cells (TICs) that do not express prostate cancer markers, such as androgen receptor or prostate specific antigen. These TICs possess stem cell characteristics and multipotency as demonstrated by in vitro sphere-formation and in vivo tumour-initiation, respectively. The cells represent an undifferentiated subtype of basal cells and can be purified from prostate tumours based on coexpression of the human pluripotent stem cell marker TRA-1-60 with CD151 and CD166. Such triple-marker-positive TICs recapitulate the original parent tumour heterogeneity in serial xeno-transplantations indicating a tumour cell hierarchy in human prostate cancer development. These TICs exhibit increased nuclear factor-κB activity. These findings are important in understanding the molecular basis of human prostate cancer.

No MeSH data available.


Related in: MedlinePlus

Triple-marker expression in other prostate tumours and patient specimens.(a) Comparative immunohistochemical characterization of DU-145, PC3, VCaP and PC-82 tumours. Scale bar, 50 μm. (b) Percent marker-positive cells maintained after sequential in vivo passages of DU-145 tumours that were initially derived from the triple-marker-positive cells. Blue, passage1 and light blue, passage2. Mean±s.d. (n=4). (c) In vitro primary sphere-formation and (d) In vivo tumour initiation. Prospectively isolated 2,500 marker-positive DU-145 tumour cells were used. Tumour size was determined as tumour volume. Mean±s.d. (n=4). Unsorted control represents single cells of tumour. (e) Immunofluorescence labelling of markers in fresh frozen human patient primary prostate tumour specimens, with 4′-6-diamidino-2-phenylindole (DAPI) counterstain. Scale bar, 100 μm. (f) Percent marker-positive cells in human patients' prostate tumour specimens. Mean±s.d. (n=6). (g) Percent marker-positive cells in a variety of epithelial tumour specimens from clinical patients. Red, tumour cells and green, patient-matched normal tissue cells. Mean±s.d. (n=3).
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f4: Triple-marker expression in other prostate tumours and patient specimens.(a) Comparative immunohistochemical characterization of DU-145, PC3, VCaP and PC-82 tumours. Scale bar, 50 μm. (b) Percent marker-positive cells maintained after sequential in vivo passages of DU-145 tumours that were initially derived from the triple-marker-positive cells. Blue, passage1 and light blue, passage2. Mean±s.d. (n=4). (c) In vitro primary sphere-formation and (d) In vivo tumour initiation. Prospectively isolated 2,500 marker-positive DU-145 tumour cells were used. Tumour size was determined as tumour volume. Mean±s.d. (n=4). Unsorted control represents single cells of tumour. (e) Immunofluorescence labelling of markers in fresh frozen human patient primary prostate tumour specimens, with 4′-6-diamidino-2-phenylindole (DAPI) counterstain. Scale bar, 100 μm. (f) Percent marker-positive cells in human patients' prostate tumour specimens. Mean±s.d. (n=6). (g) Percent marker-positive cells in a variety of epithelial tumour specimens from clinical patients. Red, tumour cells and green, patient-matched normal tissue cells. Mean±s.d. (n=3).

Mentions: The triple-marker expression and the association with tumour-initiation were consistent in additional human prostate cancer cell line-derived xenograft tumour models, namely, androgen-independent metastatic prostate cancer cell line-derived DU-145 (brain metastasis), PC3 (bone metastasis) and VCaP (vertebral metastasis) xenograft tumours, and also in another androgen-dependent and human patient-derived primary OT-xenograft tumour (PC-82; Table 2; Supplementary Fig. S5). All the resulting tumours exhibited their expected immunohistochemical characteristics (Fig. 4a) such as cytosolic localization of AR in androgen-insensitive DU-145 and PC3 tumours, or nuclear localization in androgen-sensitive VCaP and PC-82 tumours. All these tumours contained a set of triple-marker-positive cells (Table 2). Tumours derived from triple-marker-positive DU-145 tumour cells were sequentially passaged in vivo over multiple transplantation cycles (Fig. 4b). The marker-positive cells from the DU-145 tumour recapitulated the original parent tumour heterogeneity by hierarchically differeniating in vivo into both marker-positive and -negative tumour cells. Marker expression and sphere-forming/tumour-initiating abilities were also correlated in the DU-145 tumour cells, although the marker negative cells have considerably diminished sphere-forming and tumour-initiating abilities (Fig. 4c,d). We next verified the expression of each of these triple-markers in freshly resected human patient specimens from prostate tumour by immunofluorescence (Fig. 4e) and FACS (Fig. 4f) or from other epithelial tumours of breast, colon and ovary by FACS (Fig. 4g). Cells positive for expression of the TRA-1-60 and triple-marker-positive cells were enriched in other epithelial tumour specimens when compared with the respective patient-matched normal tissues.


Tumour-initiating stem-like cells in human prostate cancer exhibit increased NF-κB signalling.

Rajasekhar VK, Studer L, Gerald W, Socci ND, Scher HI - Nat Commun (2011)

Triple-marker expression in other prostate tumours and patient specimens.(a) Comparative immunohistochemical characterization of DU-145, PC3, VCaP and PC-82 tumours. Scale bar, 50 μm. (b) Percent marker-positive cells maintained after sequential in vivo passages of DU-145 tumours that were initially derived from the triple-marker-positive cells. Blue, passage1 and light blue, passage2. Mean±s.d. (n=4). (c) In vitro primary sphere-formation and (d) In vivo tumour initiation. Prospectively isolated 2,500 marker-positive DU-145 tumour cells were used. Tumour size was determined as tumour volume. Mean±s.d. (n=4). Unsorted control represents single cells of tumour. (e) Immunofluorescence labelling of markers in fresh frozen human patient primary prostate tumour specimens, with 4′-6-diamidino-2-phenylindole (DAPI) counterstain. Scale bar, 100 μm. (f) Percent marker-positive cells in human patients' prostate tumour specimens. Mean±s.d. (n=6). (g) Percent marker-positive cells in a variety of epithelial tumour specimens from clinical patients. Red, tumour cells and green, patient-matched normal tissue cells. Mean±s.d. (n=3).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3105310&req=5

f4: Triple-marker expression in other prostate tumours and patient specimens.(a) Comparative immunohistochemical characterization of DU-145, PC3, VCaP and PC-82 tumours. Scale bar, 50 μm. (b) Percent marker-positive cells maintained after sequential in vivo passages of DU-145 tumours that were initially derived from the triple-marker-positive cells. Blue, passage1 and light blue, passage2. Mean±s.d. (n=4). (c) In vitro primary sphere-formation and (d) In vivo tumour initiation. Prospectively isolated 2,500 marker-positive DU-145 tumour cells were used. Tumour size was determined as tumour volume. Mean±s.d. (n=4). Unsorted control represents single cells of tumour. (e) Immunofluorescence labelling of markers in fresh frozen human patient primary prostate tumour specimens, with 4′-6-diamidino-2-phenylindole (DAPI) counterstain. Scale bar, 100 μm. (f) Percent marker-positive cells in human patients' prostate tumour specimens. Mean±s.d. (n=6). (g) Percent marker-positive cells in a variety of epithelial tumour specimens from clinical patients. Red, tumour cells and green, patient-matched normal tissue cells. Mean±s.d. (n=3).
Mentions: The triple-marker expression and the association with tumour-initiation were consistent in additional human prostate cancer cell line-derived xenograft tumour models, namely, androgen-independent metastatic prostate cancer cell line-derived DU-145 (brain metastasis), PC3 (bone metastasis) and VCaP (vertebral metastasis) xenograft tumours, and also in another androgen-dependent and human patient-derived primary OT-xenograft tumour (PC-82; Table 2; Supplementary Fig. S5). All the resulting tumours exhibited their expected immunohistochemical characteristics (Fig. 4a) such as cytosolic localization of AR in androgen-insensitive DU-145 and PC3 tumours, or nuclear localization in androgen-sensitive VCaP and PC-82 tumours. All these tumours contained a set of triple-marker-positive cells (Table 2). Tumours derived from triple-marker-positive DU-145 tumour cells were sequentially passaged in vivo over multiple transplantation cycles (Fig. 4b). The marker-positive cells from the DU-145 tumour recapitulated the original parent tumour heterogeneity by hierarchically differeniating in vivo into both marker-positive and -negative tumour cells. Marker expression and sphere-forming/tumour-initiating abilities were also correlated in the DU-145 tumour cells, although the marker negative cells have considerably diminished sphere-forming and tumour-initiating abilities (Fig. 4c,d). We next verified the expression of each of these triple-markers in freshly resected human patient specimens from prostate tumour by immunofluorescence (Fig. 4e) and FACS (Fig. 4f) or from other epithelial tumours of breast, colon and ovary by FACS (Fig. 4g). Cells positive for expression of the TRA-1-60 and triple-marker-positive cells were enriched in other epithelial tumour specimens when compared with the respective patient-matched normal tissues.

Bottom Line: These TICs possess stem cell characteristics and multipotency as demonstrated by in vitro sphere-formation and in vivo tumour-initiation, respectively.The cells represent an undifferentiated subtype of basal cells and can be purified from prostate tumours based on coexpression of the human pluripotent stem cell marker TRA-1-60 with CD151 and CD166.These TICs exhibit increased nuclear factor-κB activity.

View Article: PubMed Central - PubMed

Affiliation: 1] Stem Cell Center and Developmental Biology Program, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA. [2] Sidney Kimmel Center for Prostate and Urologic Cancers, Department of Medicine, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA.

ABSTRACT
Androgen depletion is a key strategy for treating human prostate cancer, but the presence of hormone-independent cells escaping treatment remains a major therapeutic challenge. Here, we identify a minor subset of stem-like human prostate tumour-initiating cells (TICs) that do not express prostate cancer markers, such as androgen receptor or prostate specific antigen. These TICs possess stem cell characteristics and multipotency as demonstrated by in vitro sphere-formation and in vivo tumour-initiation, respectively. The cells represent an undifferentiated subtype of basal cells and can be purified from prostate tumours based on coexpression of the human pluripotent stem cell marker TRA-1-60 with CD151 and CD166. Such triple-marker-positive TICs recapitulate the original parent tumour heterogeneity in serial xeno-transplantations indicating a tumour cell hierarchy in human prostate cancer development. These TICs exhibit increased nuclear factor-κB activity. These findings are important in understanding the molecular basis of human prostate cancer.

No MeSH data available.


Related in: MedlinePlus