The immunohistochemical expression of stress-response protein (srp) 60 in human brain tumours: relationship of srp 60 to the other five srps, proliferating cell nuclear antigen and p53 protein.
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This study analyzed the expression of stress-response (heat-shock) protein 60 (srp 60) in a series of 158 human brain tumours. Immunohistochemical procedures were employed; cells of the human cervical cancer line HeLa S3 exposed to hyperosmolar stress served as positive controls. Deposits of reaction products were found in the cytoplasm. Approximately half of the glioblastomas multiforme (17/31), breast carcinoma metastases (6/10), and lung carcinoma metastases (5/11) as well as about one-third of the astrocytomas (5/13) and meningiomas (8/23) had tumour cells that expressed srp 60. A positive reaction for srp 60 was also seen in some medulloblastomas (2/16), primitive neuroectodermal tumours (PNETs) (2/11), schwannomas (2/21), and pituitary adenomas (2/7), but no positive reactions were observed with oligodendrogliomas and ependymomas. Compared with srp 60-negative tumours, srp 60-positive tumours coexpressed one or more stress-related proteins, among which srp 90, srp 72, srp 27, alphaB-crystallin and ubiquitin occurred with higher frequencies; a high correlation between srp 60 and the other five srps (0.88 - 0.97, p<0.01, Pearson correlation coefficient) was observed in srp 60-positive tumours. In contrast, the correlation coefficient in srp 60-negative tumours was not significant (-0.26 - 0.71). There was a tendency for the proliferating cell nuclear antigen (PCNA)-labeling index to be higher in glioblastomas, astrocytomas, medulloblastomas, PNETs, and breast and lung carcinoma metastases that expressed srp 60 than in those that did not. No significant immunohistochemical reactions of srp 60, PCNA and p53 protein were seen with sections of normal brain tissues. We conclude that primary and metastatic tumours of the brain produce srp 60 and that srp 60 in certain brain tumour cells may coexpress the other five srps. In addition, srp 60 expression might depend, in part, on proliferating potential.Immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA) and p53 protein is important, particularly for the surgical diagnosis of neoplastic disorders. An effective, simple and reproducible method was established for observing the expression of these intranuclear antigens in routinely processed, formalin-fixed paraffin-embedded sections. Dramatic improvement of the antigenicity was obtained when the deparaffinized sections were heated in a hot water bath at 90 degrees C for 120 min in 0.01 mol/L citrate buffer, pH 6.0, for PCNA and in 0.01 mol/L phosphate-buffered saline, pH 7.2, for p53 protein. These reliable pretreatments are useful for the detailed comparative analysis of the expression of PCNA and p53 protein and fine histologic architecture and for retrospective study using a large number of archival specimens.
Antigenicity
Phosphate buffered saline
Antigen retrieval
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The possibility that proliferating cell nuclear antigen (PCNA) is expressed by non-proliferating liver cells was investigated. Liver biopsies from 107 patients were investigated, which included histologically normal liver, metastatic tumour, and inflammatory lesions. PCNA was detected using immunohistochemical staining with the monoclonal antibody PC10. This was compared with the proportion of proliferating cells as assessed by immunostaining for the Ki-67 antigen using the monoclonal antibody MIB 1. Most cases of histologically normal liver showed few PC10-positive cells. PCNA-positive hepatocytes far outnumbered those positive with MIB 1 in specimens showing metastatic tumour or an inflammatory cell infiltrate. There was no relation between the degree of PCNA overexpression and the type of tumour present or the nature of the inflammatory lesion. Other cell types, including the biliary epithelium, did not show this large difference between the proportions of PC10- and MIB 1-positive cells. It is concluded that non-proliferating hepatocytes increase their levels of PCNA in a wide variety of pathological conditions. This may be mediated by cytokines released by tumour cells or inflammatory cells.
Immunostaining
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Immunohistochemical study of proliferating cells in normal or neoplastic tissues has its advantages over other methods in understanding cell kinetic information. Proliferating cell nuclear antigen (PCNA) immunolocalization in paraffin section was reported as an index for cellular proliferation and well-correlated with the histopathological grading of certain systemic malignancies.By using immunohistochemical staining of PCNA on formalin-fixed, paraffin-embedded sections, the anti-PCNA monoclonal antibody (PC10) staining scores were counted in 32 intracranial gliomas and 32 intracranial meningiomas.The PC10 scores ranged from 7.8% to 24.1% (m = 15.7 +/- 4.5%) in 12 anaplastic astrocytomas, and from 15.1% to 51.3% (m = 38.4 +/- 11.7%) in 9 glioblastomas. The difference in between was significant. The mean PC10 scores of benign meningiomas, malignant meningiomas, and hemangiopericytomas were 13.8%, 50.2%, and 50.8% respectively. The difference of PC10 scores was also significant between benign and malignant meningioma, as well as between benign meningioma and hemangiopericytoma.The PC10 scores have a good correlation with the histopathological grading of both intracranial glioma and meningioma.
Grading (engineering)
Hemangiopericytoma
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Cutaneous histiocytoma is a benign tumour known to occur in dogs only. The present study deals with a case of cutaneous histiocytoma diagnosed histopathologically in a buffalo. Immunohistochemical investigation revealed strong nuclear immunolabelling of tumour cells for p53 and c-Myc oncoproteins and proliferating cell nuclear antigen (PCNA). The neoplastic cells exhibited very weak expression of telomerase catalytic unit, hTERT. Pericellular fibronectin (FN) was seen in most of the tumour cells while scanty connective tissue stroma showed light FN immunolabelling. Observations of high indices for cell proliferation markers such as mitoses (8.03±2.19), AgNORs (9.76±2.36) and PCNA (545.20±134.00) revealed aggressive proliferative activity of the tumour cells. The study recorded the case of cutaneous histiocytoma for the first time in a buffalo. The findings of immunohistochemistry suggested the aggressive growth behaviour of the cutaneous histiocytoma, however, the absence of cellular immortality, invasiveness and motility proved its benign nature.
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SUMMARY In this study, the expression of proliferating cell nuclear antigen (PCNA) and p53 proteins was investigated in canine mammary tumours and in healthy tissues by immunohistochemistry using the PC10 anti-PCNA antibody and the DO7 and PAb240 anti-p53 antibodies. A total of 46 cases (19 benign lesions: hyperplasia, adenomas and benign mixed tumours and 27 malignant lesions: carcinomas and malign mixed tumours) were examined. No PCNA and p53 staining was evidenced in healthy mammary tissues. The frequency of nuclear PCNA positive staining was higher among malignant tumours (88.9%) than in benign lesions (52.6%) in which the staining extent was always low ( 10%) to high (> 50%) in 75% of the positive malignant tumours, mainly in solid adenocarcinomas and in papillar adenocarcinomas at a lesser extent. Whereas the DO7 antibody failed to detect any p53 accumulation, nuclear and cytoplasm p53 staining was evidenced using the Pab240 antibody in 15.8% and 48.1% of benign and malignant tumours respectively. The p53 staining remained weak (< 10%) in benign lesions and was more intense in malignant tumours mainly in adenocarcinomas (66.7% of positive solid adenocarcinomas exhibited more than 10% of positive cells). Moreover, p53 positive tumours were also PCNA positive except for one case. These results show that PCNA and p53 may be useful markers for evaluating malignancy in canine mammary tumours.
Atypical Hyperplasia
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Aim To study expression and significance of proliferating cell nuclear antigen(PCNA) and vimentin in renal tissues of the patients with epidemic hemorrhagic fever(EHF). Methods 17 autopsy renal tissues from the patients died of EHF were examined by immunohistochemical staining(PAP). Results The results showed that positive rate of PCNA expression in 17 autopsy renal tissues was 6471% ; proliferative index of collecting tubes was significantly higher than that of distal convoluted tubules(P 0001); Vimentin expression of epithelial cells in distal convoluted tubules and collecting tubules from the EHF patients at hypotensive shock stage were strongly positive,PCNA expression at the same position was also positive,and positive intensity of the PCNA expression was consistent with severity of local lesions. Conclusion There are regeneration,proliferation and differentiation of injured renal tubular epithelial cells at early stage of the EHF patients with acute renal failure. These changes of the renal tubular epithelial cells are of great significance during restoration of injured kidneys.
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Objectives: To assess the immunohistochemical expression of Ki-67 and Proliferating Cell Nuclear Antigen (PCNA) as proliferative markers to study proliferative activity and CD34 as an endothelial cell marker in order to study vascular proliferation in astrocytomas in correlation with some clinicopathological parameters (age, gender, site of the tumor, and tumor grade).Methods: A retrospective study wherein a total of 51 formalinfixed paraffin-embedded brain astrocytoma excisional biopsies covering the period of June 2009 to February 2011 were retrieved from the archival materials of the Specialized Surgical Hospital in Medical City in Baghdad, Iraq.The histopathological diagnosis had been revised and all cases were stained by immunohistochemical technique with Ki-67, PCNA, and CD34 tumor markers.Values were considered statistically significant when p<0.05.Results: Fibrillary astrocytoma (WHO grade II) was found to be the most common type among astrocytic tumors with the peak age incidence of astrocytomas found in the second and fifth decades of life, and a slight male predominance had been identified.There was a significant correlation between the age of the patients and the grade of the tumor, Ki-67 and PCNA labeling indices, and microvessel density (MVD) detected by CD34 (p<0.05).There was a highly significant correlation between Ki-67 and PCNA labeling indices in astrocytomas (p<0.001).Conclusion: A significant correlation was found between Ki-67, PCNA labeling indices, and MVD (microvessel density) detected by CD34, and between the clinicopathological variables of astrocytomas (age and grade of tumor).Hence, Ki-67 and PCNA, as markers for proliferation, and MVD as a marker of angiogenesis, could be used as ancillary methods in the differentiation of borderline grades of astrocytomas.
Ki-67
Anaplastic astrocytoma
Proliferation Marker
Microvessel
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