• Characterization of the Hoechst 33342 side population from normal and malignant human renal epithelial cells.

      Addla, Sanjai K; Brown, Michael D; Hart, Claire A; Ramani, Vijay A C; Clarke, Noel W; Genito-Urinary Cancer Research Group, School of Cancer and Imaging Sciences, Paterson Institute for Cancer Research, University of Manchester, Wilmslow Road, Manchester, UK. (2008-09)
      The fundamental changes which predispose for renal cell carcinoma (RCC) are poorly characterized. It is hypothesized that "cancer stem cells" may be influential in carcinogenesis, and the epithelial side population (SP) is enriched for stemlike cells in other epithelial cancers. In this study, we have isolated and characterized the SP and non-SP (NSP) populations from normal (NK) and malignant (RCC) human kidney tissue. NK specimens were taken from patients undergoing non-renal cancer surgery and paired malignant and macroscopically normal tissue samples were taken from patients undergoing surgery for RCC. The Hoechst 33342 dye efflux technique was used to isolate epithelial SP and NSP from normal and malignant human renal tissue. Cellular subpopulations were phenotyped for lineage, cell cycle, and putative stem cell markers, and functionally characterized using in vitro colony-forming and proliferation assays. The SP constituted 3.8 +/- 0.4 and 5.9 +/- 0.9% of epithelial cells in NK and RCC, respectively, of which 14.1 +/- 3.5 and 13.2 +/- 3.6% were shown to be in G(0). SP cells demonstrated greater proliferative potential in colony-forming efficiency, long-term culture, and spheroids assays and were shown to be maintained upon tissue culture passage. We have shown that the renal SP is enriched for quiescent cells, with a high proliferative capacity and stemlike properties. The population is, however, heterogeneous, confirming that the terms "SP cell" and "stem cell" cannot be used interchangeably.
    • Histological outcome of delayed orchidectomy after primary chemotherapy for metastatic germ cell tumour of the testis.

      Ramani, Vijay A C; Grey, Benjamin R; Addla, Sanjai K; Dunham, Mark P; Sangar, Vijay K; Clarke, Noel W; Christie Hospital NHS Foundation Trust, Manchester, UK. (2008-04)
      AIMS: To identify the incidence of viable local tumour in the testis of patients undergoing delayed orchidectomy after initial presentation with advanced germ cell tumour (GCT) treated by primary chemotherapy. PATIENTS AND METHODS: Thirty-three patients presenting with advanced metastatic GCT were reviewed. The median age at presentation was 34 years. All received chemotherapy without previous orchidectomy. The decision to initiate chemotherapy without orchidectomy was based on a heavy tumour load and the patient's condition at initial presentation. A histological diagnosis was available from a biopsy of metastases in 23 patients; treatment in the remaining 10 patients was initiated after diagnosis based on a combination of elevated serum tumour markers, testicular findings and the presence of a retroperitoneal mass. RESULTS: Seminomatous GCT (SGCT) was diagnosed in 13 patients, non-seminomatous GCT (NSGCT) in 17 patients and mixed GCT (MGCT) in the remaining three patients. Bleomycin/etoposide/cisplatin-based chemotherapy was the principle regimen. After initial chemotherapy, all patients with pure SGCT had only scar tissue in the orchidectomy specimen, with no residual tumour. Nine of 17 patients (52.9%) with NSGCT had viable tumour remaining in the orchidectomy specimen. All three cases of MGCT had persistent viable invasive seminoma. Twenty-seven patients (81.8%) were recurrence free and alive after a median of 49 months of follow-up. CONCLUSIONS: Thirty-six per cent of patients had residual tumour locally in the testis after primary chemotherapy for metastatic GCT of the testis. However, in the cases with pure seminomatous disease, there was no residual tumour present. It may not be necessary to undertake delayed orchidectomy in these patients.