Craig N. Robson

CD133, a novel marker for human prostatic epithelial stem cells

Stem cells are clonogenic cells with self-renewal and differentiation properties, which may represent a major target for genetic damage leading to prostate cancer and benign prostatic hyperplasia. Stem cells remain poorly characterised because of the absence of specific molecular markers that permit us to distinguish them from their progeny, the transit amplifying cells, which have a more restricted proliferative potential. Human CD133 antigen, also known as AC133, was recently identified as a haematopoietic stem cell marker. Here we show that a small population (approximately 1%) of human prostate basal cells express the cell surface marker CD133 and are restricted to the α2β1hi population, previously shown to be a marker of stem cells in prostate epithelia (Collins, A. T., Habib, F. K., Maitland, N. J. and Neal, D. E. (2001). J. Cell Sci. 114, 3865-3872). α2β1hi/CD133+ cells exhibit two important attributes of epithelial stem cells: they possess a high in vitro proliferative potential and can reconstitute prostatic-like acini in immunocompromised male nude mice.

Expression of Bcl-2, Bax, and p53 in high-grade prostatic intraepithelial neoplasia and localized prostate cancer: relationship with apoptosis and proliferation

Apoptosis‐regulating genes have been shown to be important in the biology of prostate cancer. The aim of this study was to examine and correlate the expression of the apoptosis‐regulating genes bcl‐2, bax, and p53 with the frequency of apoptosis and rate of proliferation in benign prostatic epithelium (BP), prostate cancer, and high‐grade prostatic intraepithelial neoplasia (HGPIN), which is currently considered the most likely precursor of prostate cancer.

Localization and quantification of mRNA for matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in human benign and malignant prostatic tissue.

The family of matrix metalloproteinases (MMPs) has been shown to be involved in proteolytic degradation of the extracellular matrix, which is an essential step in tumor invasion and metastasis. MMPs are tightly regulated by the levels of active enzymes and their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). MMP‐2 and its ratio to TIMP‐2 have been associated with tumor recurrence and progression in a number of human malignancies.

Fibroblast growth factor 17 is over-expressed in human prostate cancer

Over‐expression of fibroblast growth factor 8 (FGF8) in human prostate cancer is associated with clinically aggressive disease. Among different members of the FGF family, FGF17 and FGF8 share high sequence homology and have similar patterns of expression during embryogenesis. In this study, the clinical significance of FGF17 expression and its in vitro function in prostate cancer cells were tested. Forty resected prostate specimens from patients with benign prostatic hyperplasia (BPH, n = 12) and prostate cancer (CaP, n = 28; Gleason sum scores 3–10) were studied using semi‐quantitative RT‐PCR. In addition, 85 cases of CaP (Gleason sum scores 5–9) and 20 cases of BPH were examined using immunohistochemistry and findings were correlated with clinical parameters. In vitro experiments using prostate cancer cell lines examined the functional significance of FGF17 in prostate cancer. These studies revealed a significant linear correlation between increasing Gleason sum scores and FGF17 expression using both immunohistochemistry (p < 0.0001, ρ = 0.99) and RT‐PCR (p = 0.008, ρ = 0.99). Immunohistochemistry demonstrated upregulation of FGF17 in CaP compared with BPH (p < 0.0001) and, when comparing high‐grade CaP (Gleason sum score 7–10) with BPH, RT‐PCR showed a fourfold upregulation of FGF17 mRNA expression (p < 0.0001). Men with tumours displaying high levels of FGF17 expression had a worse outcome on survival analysis (p = 0.044) and a higher risk of progression with metastases (p < 0.0001). Proliferation assays showed low‐dose recombinant (r) FGF17 (1 ng/ml) to be a more potent mitogen than rFGF1 and rFGF8 in prostate cancer cell lines (LNCaP, DU145, and PC3M) (p < 0.001). Furthermore, FGF8 was shown to induce expression of FGF17 in these cell lines. These data support a role for FGF17, and a model of co‐expression of multiple FGFs, with FGF17 as a potential mediator of FGF8 function, in human prostate carcinogenesis. Copyright

Androgen regulation of ornithine decarboxylase in human prostatic cells identified using differential display

Androgens are essential for normal prostate physiology and have a permissive role in the development and progression of prostate cancer. Using the mRNA differential display technique, ornithine decarboxylase (ODC) was identified to be up‐regulated by androgens in human prostatic LNCaP cells. On Northern analysis, the induction of ODC expression by 10 nM androgen was rapid and continued up to 48 h exposure with a maximum 6.3‐fold up‐regulation. The anti‐androgen Casodex inhibited the androgen‐induced up‐regulation of ODC, whereas the protein synthesis inhibitor cycloheximide did not. Together these data suggest that regulation is mediated through the androgen receptor protein and does require secondary protein synthesis, respectively. The kinetics of induction of ODC were almost identical to those of prostate specific antigen. Taken together these data suggest that ODC is directly regulated by androgens in LNCaP cells.

Fibroblast growth factor receptor-2 mutation analysis in human prostate cancer

Objective To assess whether mutations in the hot‐spots of the fibroblast growth factor (FGF) receptor‐2 gene (FGFR2, exons encoding the IIIa, IIIb, IIIc and transmembrane domain, TMD) are associated with the development of prostate cancer, as the IIIb variant is the specific receptor for FGF7/KGF, an androgen‐inducible paracrine factor regulating prostatic growth.

Effects of a new 5α reductase inhibitor (epristeride) on human prostate cell cultures

Inhibitors of 5α reductase (5αR), the enzyme that converts testosterone to dihydrotestosterone (DHT), have been shown to retard the growth of hyperplastic prostates. This study evaluates the effects of the 5αR inhibitor, epristeride, on cultured stromal and epithelial cells from benign, hyperplastic adult prostates.

A novel laparoscopically-deployed device for renal hypothermia: cooling efficacy in a porcine model

ã 2007 THE AUTHORS JOURNAL COMPILATION ã 2007 BJU INTERNATIONAL | 99, SUPPLEMENT 4, 16-17 16 Tuesday 19 June 2007 18 Poster Session 1 1130–1230 Lomond Auditorium Bladder Cancer Chairmen: Mr David Gillatt and Mr Adam Jones Posters P01 – P06 19 Poster Session 2 1330–1430 Boisdale Room Stones Chairmen: Mr Adrian Joyce and Mr Daron Smith Posters P7 – P15 22 Poster Session 3 1430–1530 Boisdale Room Bladder Cancer – Diagnosis Chairmen: Mr John Anderson and Mr Derek Rosario Posters P16 – P22 24 Poster Session 4 1530–1630 Boisdale Room Governance – Clinical Chairmen: Mr Derek Fawcett and Mr Kieran O’Flynn Posters P23 – P30

The single LXXLL motif in Tip60 mediates interaction with class I nuclear hormone receptors

Background and objectives Ependymomas are the third most common CNS tumours in childhood, and account for 9–12% of CNS neoplasms in all age groups. However, the prognosis for cases not completely excised is poor and the underlying biology of their development and progression is poorly understood. The few studies published to date have suggested that specific chromosomal abnormalities may be associated with the development of a significant proportion of these tumours. We set out to screen a large series of intracranial and spinal ependymomas for genetic imbalances, and to correlate these with histology and clinical behaviour. Methods Comparative genomic hybridisation (CGH) was used to detect chromosome imbalances on 86 ependymomas from 77 patients, of which 22 were children under 16, treated at one of three UK centres (Newcastle, Nottingham, Southampton). Cases were first analysed without reference to histology or clinical features, and were then divided up according to anatomical location, histology and age at presentation. Results Chromosomal imbalances were detected in tumours from 63/77 patients (82%). The majority involved entire chromosomes or chromosome arms, many showing patterns of gains suggestive of intermediate ploidy. Of previously reported abnormalities in ependymoma, the most frequent findings were gain of 1 q, seen in 13 cases (17%), and loss of 22 in 20 (26%). Whereas 1 q gain was seen mainly in posterior fossa tumours and was restricted to those with classic and anaplastic histology, loss of 22 was rarely observed in tumours from this location and their histology was predominantly classic or myxopapillary. In contrast to previous studies, loss of 6q was found in only 6 cases (8%) and in only one child. Out of 7 tumours biopsied at presentation and relapse, 4 revealed imbalances and 3 of these demonstrated progression of abnormalities at relapse. Conclusions Chromosomal imbalance is common in ependymoma and patterns of abnormality are emerging that are associated with histology or location. Further studies are needed to establish the prognostic significance of these abnormalities.Aims Endothelin, the most potent vasoconstrictor known has been implicated development and spread of malignancy. In this study, we assessed the produc endothelin-1 (ET-1) and its precursor big endothelin (big ET-1) by human cancer cel Methods Ten human cancer cell lines were cultured (lung n = 4, colorectal n = 3, gastro-oesophageal n = 2, pancreatic n = 1). The culture media were replaced wi fresh media after the cells attained confluence. After 48 hours, the conditioned were batch analysed for ET-1 and big ET-1 by using a sandwich enzyme l immunoassay (ELISA) (Biomedica, Austria). To elucidate the action of endot converting enzyme (ECE), big ET-1 was added to one of the oesophageal canc lines after they attained confluence. Similarly, the media were analysed fo presence of ET-1 and big ET-1 Results All the ten cancer cell lines produced ET-1 and nine of ten cancer cell produced big ET-1. ET-1 and big ET-1 were not produced in equimolar amounts ratio of ET-1 to big ET-1 was 0.56–11.88 (range). All three colorectal cancer cell and four of the lung cancer cell lines produced both ET-1 and Big Et-1. Interest the oesophageal cancer cell line that produced high concentrations of ET-1 d produce any measurable big ET-1. Addition of Big ET-1 into this cell line mediu assess the action of ECE, measuring ET-1 and big ET-1 after 48 hours resu complete cleavage of big ET-1 and there was no measurable big ET-1 in the me

Expression of Tip60, an androgen receptor co-activator, in prostate cancer

Background and objectives Ependymomas are the third most common CNS tumours in childhood, and account for 9–12% of CNS neoplasms in all age groups. However, the prognosis for cases not completely excised is poor and the underlying biology of their development and progression is poorly understood. The few studies published to date have suggested that specific chromosomal abnormalities may be associated with the development of a significant proportion of these tumours. We set out to screen a large series of intracranial and spinal ependymomas for genetic imbalances, and to correlate these with histology and clinical behaviour. Methods Comparative genomic hybridisation (CGH) was used to detect chromosome imbalances on 86 ependymomas from 77 patients, of which 22 were children under 16, treated at one of three UK centres (Newcastle, Nottingham, Southampton). Cases were first analysed without reference to histology or clinical features, and were then divided up according to anatomical location, histology and age at presentation. Results Chromosomal imbalances were detected in tumours from 63/77 patients (82%). The majority involved entire chromosomes or chromosome arms, many showing patterns of gains suggestive of intermediate ploidy. Of previously reported abnormalities in ependymoma, the most frequent findings were gain of 1 q, seen in 13 cases (17%), and loss of 22 in 20 (26%). Whereas 1 q gain was seen mainly in posterior fossa tumours and was restricted to those with classic and anaplastic histology, loss of 22 was rarely observed in tumours from this location and their histology was predominantly classic or myxopapillary. In contrast to previous studies, loss of 6q was found in only 6 cases (8%) and in only one child. Out of 7 tumours biopsied at presentation and relapse, 4 revealed imbalances and 3 of these demonstrated progression of abnormalities at relapse. Conclusions Chromosomal imbalance is common in ependymoma and patterns of abnormality are emerging that are associated with histology or location. Further studies are needed to establish the prognostic significance of these abnormalities.Aims Endothelin, the most potent vasoconstrictor known has been implicated development and spread of malignancy. In this study, we assessed the produc endothelin-1 (ET-1) and its precursor big endothelin (big ET-1) by human cancer cel Methods Ten human cancer cell lines were cultured (lung n = 4, colorectal n = 3, gastro-oesophageal n = 2, pancreatic n = 1). The culture media were replaced wi fresh media after the cells attained confluence. After 48 hours, the conditioned were batch analysed for ET-1 and big ET-1 by using a sandwich enzyme l immunoassay (ELISA) (Biomedica, Austria). To elucidate the action of endot converting enzyme (ECE), big ET-1 was added to one of the oesophageal canc lines after they attained confluence. Similarly, the media were analysed fo presence of ET-1 and big ET-1 Results All the ten cancer cell lines produced ET-1 and nine of ten cancer cell produced big ET-1. ET-1 and big ET-1 were not produced in equimolar amounts ratio of ET-1 to big ET-1 was 0.56–11.88 (range). All three colorectal cancer cell and four of the lung cancer cell lines produced both ET-1 and Big Et-1. Interest the oesophageal cancer cell line that produced high concentrations of ET-1 d produce any measurable big ET-1. Addition of Big ET-1 into this cell line mediu assess the action of ECE, measuring ET-1 and big ET-1 after 48 hours resu complete cleavage of big ET-1 and there was no measurable big ET-1 in the me