Pamela J. Russell

Ectopic hormone production by a prostatic small cell carcinoma xenograft line

The xenograft line, UCRU-PR-2, has been characterized further. Established from a primary human undifferentiated small cell carcinoma of the prostate, it has been maintained as a stable xenograft line in nude mice and is currently in passage 9. The tumor has maintained the features of small cell undifferentiated carcinoma but shows epithelial as well as neuroendocrine characteristics. In this paper, we describe synthesis and secretion of peptide hormones, ACTH, beta-endorphin and somatostatin in vivo and ACTH and beta-endorphin in vitro by the tumor, UCRU-PR-2. This suggests that the gene for proopiomelanocortin is expressed and that processing of the molecule occurs. This line may yield insights into the histogenesis of the subtypes of prostate cancer, and also aid studies of regulation of ectopic hormone production.

Detection of malignant cells in voided urine from patients with bladder cancer, a novel monoclonal assay.

A simple assay is described for detecting malignant cells in the voided urine from patients with transitional cell carcinoma of the bladder. Agarose-embedded urothelial cells from 24 biopsy-proven cancer patients and 10 controls were stained for surface immunofluorescence with four monoclonal antibodies reactive with human bladder cancer and three monoclonals reactive with blood group A. Reactivity was assessed by fluorescence microscopy. One antibody, BLCA-8 appeared to have particular diagnostic utility. Thus, 24.3 +/- 5.8 percent of outer layer and 27.0 +/- 4.6 percent of inner layer urothelial cells reacted with BLCA-8 in patient samples, compared to 2.9 +/- 1.0 and 0.8 +/- 0.5 percent of similar cells from control urines. BLCA-8 antigen expression was found to be relatively stable even after prolonged exposure to urine. In a comparison with conventional cytology, samples from 4/8 patients were considered positive by standard methods, whereas, 8/8 were BLCA-8 positive. This new technique may thus be a useful adjunct to conventional methods.

Prostate cancer methods and protocols

Epidemiological Investigation of Prostate Cancer Graham G. Giles Human Prostate Cancer Cell Lines Pamela J. Russell and Elizabeth A. Kingsley Growth of Prostatic Epithelial and Stromal Cells In Vitro Donna M. Peehl Prostate Epithelial Stem Cell Isolation and Culture David L. Hudson and John R. W. Masters Generation of Immortal Human Prostate Cell Lines for the Study of Prostate Cancer Johng S. Rhim Spheroids of Prostate Tumor Cell Lines George Sgouros, Wei-Hong Yang, and Richard Enmon Animal Models of Prostate Cancer Pamela J. Russell and Dale J. Voeks Transgenic Mouse Models for Prostate Cancer: Identification of an Androgen-Dependent Promoter and Creation and Characterization of the Long Probasin Promoter-Large T Antigen (LPB-Tag) Model Susan Kasper, William Tu, Richard L. Roberts, and Scott B. Shappell In Vivo Models of Human Prostate Cancer Bone Metastasis Julie M. Brown Effects of Fixation on Tissues Elin Mortensen and Julie M. Brown Background, Methods, and Protocols for the Histopathological Diagnosis of Prostate Carcinoma Warick Delprado Realizing the Potential of Ejaculate/Seminal Fluid in Detecting and Predicting Natural History R. A. Gardiner, Michelle Burger, Judith A. Clements, and Martin F. Lavin Bisulfite Methylation Analysis of Tumor Suppressor Genes in Prostate Cancer from Fresh and Archival Tissue Samples Susan J. Clark, Douglas S. Millar, and Peter Molloy Production and Characterization of Antipeptide Kallikrein 4 Antibodies: Use of Computer Modeling to Design Peptides Specific to Kallikrein 4 Tracey J. Harvey, Ying Dong, Loan Bui, Russell Jarrott, Terry Walsh, and Judith A. Clements The Androgen Receptor CAG Repeat and Prostate Cancer Risk Peter E. Clark, Ryan A. Irvine, and Gerhard A. Coetzee Studies on Androgen Receptor Mutations and Amplification in Human Prostate Cancer Zoran Culig, Alfred Hobisch, Martin Erdel, Georg Bartsch, and Helmut Klocker Proteomics in the Analysis of Prostate Cancer Soren Naaby-Hansen, Kohji Nagano, Piers Gaffney, John R. W. Masters, and Rainer Cramer Application of Gene Microarrays in the Study of Prostate Cancer Colleen C. Nelson, Douglas Hoffart, Martin E. Gleave, and Paul S. Rennie Enhancer Trap Method Using a Green Fluorescent Protein Reporter Plasmid for Cloning Tissue-Specific Enhancers Active in Prostate Cells Fujiko Watt and Peter Molloy Targeted Alpha Therapy of Prostate Cancer Barry J. Allen, Yong Li, Syed M. A. Rizvi, and Pamela J. Russell Phenotypic and Functional Differences of Dendritic Cells Generated Under Different In Vitro Conditions Stephanie E. B. McArdle, Selman A. Ali, Geng Li, Shahid Mian, and Robert C. Rees Flavonoid Compounds in the Prevention and Treatment of Prostate Cancer Graham E. Kelly and Alan J. Husband Index

Characterization of a New Human Bladder Cancer Cell Line, Ucru-BL-28

A new human bladder cancer cell line, UCRU-BL-28 has been established and characterized from a relapsed, cisplatin resistant, grade II, stage T4 tumor. This line is tumorigenic in nude mice and reflects the pathology of the original tumor. The morphology, the expression of tumor-associated antigens and EGF receptors, and the ability to grow both in an anchorage independent manner and in the absence of serum is explored. The BL-28 line has 71-74XXY chromosomes, with del 5q, der(9) and i(19q). Further studies on the molecular basis of bladder cancer, chemosensitivity to cisplatin, growth factor production and tissue invasion are under way.

Karyotypic analysis of a heterogeneous human transitional cell carcinoma of the bladder

The UCRU-BL-17 (BL-17) series of xenografts, tissue culture sublines, and cloned cell lines (Fig. 1) shows a range of heterogeneous growth characteristics both in vitro and in vivo (Table 1) and represents a model of human bladder cancer heterogeneity. Cytogenetic analysis was undertaken to determine if specific chromosome changes correlated with particular aspects of the heterogeneous phenotypes. The BL-17 sublines and cloned cell lines shared many common chromosome abnormalities. Indeed, the cloned cell lines showed nearly identical karyotypes despite their marked differences in growth characteristics. Karyotypic evolution with passage through the nude mice was apparent, however. This evolution occurred at the specific chromosome regions of 1p12, 3cen-3p21, and 6cen-6q21. Whether the heterogeneity of karyotype between the BL-17 cell lines resulted from the existence of multiple clones in the original patient tumor or from karyotypic instability through passage in nude mice is uncertain, but in either case the specificity of karyotypic evolution observed suggests that 1p12, 3cen-3p21, and 6cen-6q21 are hotspots for rearrangement in the BL-17 tumor. No specific correlations between chromosome abnormalities and biologic characteristics could be made, but several unique karyotypic features arose in the progression to two of the sublines, BL-17/23 alpha and BL-17/0/X2A, coinciding with a loss of anchorage-independent growth by BL-17/23 alpha and a change in growth in vivo from a solid tumor to a fluid-filled tumor by BL-17/0/X2A.

Analysis of Expressed N-ras Mutations in Human Melanoma Short-term Cell Lines with Allele Specific Restriction Analysis Induced by the Polymerase Chain Reaction

Mutations in ras genes have been found in the DNA of numerous cancer types including melanomas, but the expression of these mutations in melanomas has not yet been addressed. We have used the polymerase chain reaction (PCR) and allele-specific restriction analysis (ASRA) to determine the frequency of expressed N-ras mutations on 25 short-term melanoma tissue culture samples. N-ras cDNA generated using reverse transcriptase from whole cells was used as the PCR template. 14 secondary melanoma cultures that varied in differentiation patterns were analysed. Only 2 were found to express N-ras mutations; in both, the mutation was localised to one of the first two positions of the 61st codon of N-ras. These tumour lines, KMI-M8412a and KMI-M8412b, were established from separate tumour deposits in the same patient. Codons 12 and 13 were found to be free of mutations in all of the lines studied. 8 primary melanomas and 3 unclassified skin lesions were also analysed and found free of N-ras mutations. These results suggest that N-ras may not play such an important role in melanoma tumorigenesis as is speculated by others.

Antigenic Variation and Macrophage Infiltration of Human Bladder-Tumors Xenografted Into Nude-Mice

The presence of macrophages both within and around human bladder transitional cell carcinomas xenografted into nude mice has been examined using Immunocytochemical staining. Nine different xenograft lines derived from bladder tumors of eight patients were stained for F4/80, MHC II and PGP‐1 labelled cells. The results revealed considerable heterogeneity both within and between tumors. All of the tumors generated some macrophage response at the tumor periphery, and this was marked in two instances, UCRU‐BL‐13, passage 9 and UCRU‐BL‐17, passage 2. In only two tumors was there substantial penetration of the tumor epithelium by macrophages, though infiltrating la+, F4/80+, PGP‐1+ cells were sometimes seen spreading along the base of the epithelial sheets. Three of the tumors were characterized by the presence of L3T4+ T lymphocytes at the invasion front. The presence of macrophages or lymphocytes either at the invasion front or within the tumors did not correlate with the pathological grade of the tumors. PGP‐1 monoclonal antibodies also stained granulocytes, which were present within the tumor mass in UCRU‐BL‐15, passage 2, possibly reflecting the production by the tumor of G‐CSF. In addition, the PGP‐1 antibodies stained some of the bladder tumors cells themselves, and, in some cases, the interstitial structures within the tumors. The significance of this staining is not yet understood. The xenografted tumors will provide a useful model to examine the constraints on tumor therapy using macrophage activating stimuli.