ABSTRACT
Claspin is a nuclear protein involved in DNA replication and the DNA damage response. Its structural and functional properties suggest that it may represent a potentially useful proliferation marker. To this end, a monoclonal antibody was generated and the expression of claspin was investigated in normal fibroblasts and various cancer cell lines, as well as in tumour and normal tissues from patients with primary epithelial carcinomas. Immunoblotting analysis confirmed the specificity of the antibody, while immunohistochemistry demonstrated its applicability in archival material. In normal cells and tissues, claspin expression was weak, whereas increased levels were observed in cancer cell lines and tumour specimens. Claspin staining correlated strongly with Ki67 staining in both normal (p < 0.001) and tumour tissues (p < 0.001). However, the labelling index (LI) of claspin was consistently lower than that of Ki67, suggesting that claspin expression may be limited to a narrower part of the cell cycle. Co-localization assays with cyclin A and cell synchronization experiments indicated that claspin expression coincides with the S phase. Interestingly, the relative increase of the claspin LI in tumour samples compared with normal tissues was significantly higher (14-fold) than that of the Ki67 LI (five-fold), suggesting that claspin may be a more sensitive marker of aberrant proliferation.
Subject(s)
Adaptor Proteins, Signal Transducing/analysis , Biomarkers, Tumor/analysis , Neoplasms/pathology , S Phase , Antibodies, Monoclonal/isolation & purification , Blotting, Western/methods , Carcinoma/chemistry , Carcinoma/pathology , Case-Control Studies , Cell Line , Cell Proliferation , Colorectal Neoplasms/chemistry , Colorectal Neoplasms/pathology , Cyclin A/analysis , DNA Repair , DNA Replication , Fibroblasts/chemistry , Fluorescent Antibody Technique, Indirect/methods , Humans , Immunohistochemistry/methods , Ki-67 Antigen/pharmacology , Lung Neoplasms/chemistry , Lung Neoplasms/pathology , Neoplasms/chemistry , Osteosarcoma/chemistry , Osteosarcoma/pathology , Statistics, NonparametricABSTRACT
In this study, the expression of epidermal growth factor receptor (EGFr) and c-erb-B-2 was evaluated in 35 formalin-fixed, paraffin-embedded cases of transitional cell carcinomas of the urinary bladder (TCCs). EGFr and c-erb-B-2 expression was assessed with immunohistochemistry, and molecular analysis of the respective genes was performed with the differential polymerase chain reaction. The results were statistically analyzed in relationship to histologic grade, stage, and clinical outcome. Strong cytoplasmic expression of the EGFr using the polyclonal antibody Ab-4 was found in three (25%) Grade 2 and eight (67%) Grade 3 TCCs (P < 0.01). Two Grade 2 (17%) and 11 (92%) Grade 3 TCCs strongly expressed the anti-c-erb-B-2 Ab-3 antibody. None of Ta/T1 TCCs cases showed strong expression against EGFr, in contrast to c-erb-B-2 where two cases (18%) were found to express an intense immunosignal. Eleven (85%) T2/T3 cases showed strong positivity either against EGFr or c-erb-B-2 (P < 0.001). Eleven patients died within 12 months after the diagnosis, and all of them showed strong expression of EGFr and c-erb-B-2. Amplification of EGFr and c-erb-B-2 genes was identified in one (3%) and four (11%) TCCs cases, respectively. All patients with amplified genes died within 12 months from the date of diagnosis. Our results indicate that simultaneous expression of EGFr and c-erb-B-2 occurs in TCCs, is related to the histologic grade and the stage of the disease, and denotes aggressive biologic behavior.
Subject(s)
Carcinoma, Transitional Cell/chemistry , ErbB Receptors/analysis , Genes, erbB-2/genetics , Receptor, ErbB-2/analysis , Urinary Bladder Neoplasms/chemistry , Aged , Aged, 80 and over , Carcinoma, Transitional Cell/genetics , Carcinoma, Transitional Cell/pathology , ErbB Receptors/genetics , Follow-Up Studies , Gene Amplification , Greece , Humans , Immunohistochemistry , Middle Aged , Neoplasm Invasiveness , Polymerase Chain Reaction , Receptor, ErbB-2/genetics , Urinary Bladder Neoplasms/genetics , Urinary Bladder Neoplasms/pathologyABSTRACT
Three cases of vesicouterine fistuli secondary to cesarean section are reported. The first 2 cases presented as vaginal urinary incontinence and cyclic hematuria, whereas the last case presented as bloody urinary loss from the vagina in the immediate postoperative period. The treatment was surgical in the former 2 cases and conservative in the latter case. The literature is briefly reviewed, and the treatment options are discussed.
