Clonal analysis by PCR and RFLP in breast cancer and precancerous lesions. Preliminary data.

In order to investigate the clonality in various benign, precancerous and malignant breast lesions, we analyzed small DNA samples from paraffin sections of various breast lesions by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) of the X-chromosome-linked phosphoglycerokinase (PGK) gene. DNA from the lesion as well as from the adjacent normal tissue were analyzed in each case. Two restriction endonucleases. Hpa-II, a methylation sensitive restriction enzyme and Bst XI, a restriction endonuclease, which recognizes this polymorphic site on the PGK gene, were used. Of 19 cases, one case of in situ ductal Ca (DCIS), two cases of invasive ductal Ca and one case of multifocal ductal Ca were shown to be monoclonal. Two cases of intraductal papillomas were found to be polyclonal. One case of atypical ductal hyperplasia showed no conclusive findings. Additionally DNA was not yielded from 3 cases and other 7 cases were not informative, derived from females homozygotes. Further study of a larger number of cases is in progress.

p53 DO-1 immunohistochemical overexpression in premalignant and malignant cervical lesions.

One hundred-fourteen cervical lesions, including paraffin sections from benign, premalignant and malignant cervical tissue specimens were examined immunohistochemically for overexpression of p53 protein, using the p53 DO-1 monoclonal antibody (MoAb). p53 overexpression was observed in 66% of premalignant cervical lesions and in 90% of invasive squamous cell carcinomas. The difference in p53 positive lesions of premalignant and malignant cervical lesions was statistically very significant (p < 0.001) and the difference between CIN1 and CIN2 and CIN3 statistically significant (p < 0.01). Our results indicate that p53 overexpression may be involved at an early stage in cervical carcinogenesis.

Topographical immunohistochemical expression of bcl-2 protein in human liver lesions.

Immunolocalization of the bcl-2 protein was investigated in 60 hepatocellular carcinomas, 10 cholangiocarcinomas. 15 metastatic adenocarcinomas as well as in 37 non-neoplastic liver lesions. The three-step immunoperoxidase method was performed in archival, routinely processed material. bcl-2 protein was not identified either in neoplastic, dysplastic or normal hepatocytes, whereas it was observed in bile ductules and small bile duct epithelia, but not in the epithelium lining large bile ducts. All cases of cholangiocarcinoma and 60% of metastatic adenocarcinomas were bcl-2 positive. bcl-2 appears to be an additional marker in distinguishing hepatocellular carcinoma from cholangiocarcinoma or metastatic adenocarcinoma. Also, bcl-2 does not seem to be involved in human liver hepatocyte survival.

p53 expression in hepatocellular carcinoma in Greece. Correlation with epidemiological and histopathological data.

Localization of p53 oncoprotein was investigated in 60 hepatocellular carcinomas (HCCs) from patients resident in the Northwest and Central Greece. The streptavidin-biotin complex immunoperoxidase method was performed in archival formalin-fixed, paraffin-embedded material, using the monoclonal antibody DO-1. The aim of our study was to correlate p53 expression with histological and epidemiological data. p53 overexpression in patients with serological hepatitis B or C was greater (47%) as compared to that observed in patients without these markers (p < 0.01). Morphologically normal liver tissue (NLT) and liver cell dysplasia (LCD) was recognized adjacent to HCC developing on non-alcoholic cirrhotic livers in patients with "NonA, NonB hepatitis" from between 1975-1986. NLT and LCD and p53 oncoprotein was expressed in 10% of the cases. No relationship was observed between p53 expression and tumor histological grade, patients' age and sex. These results suggest that in Northwest and Central Greece, p53 oncosupressor gene may be involved in some HCCs; it may be associated with viral chronic infection disease (HBV or HCV), and as yet with uncharacterized viruses which remain to be determined.

p53 and c-jun expression in urinary bladder transitional cell carcinoma: correlation with proliferating cell nuclear antigen (PCNA) histological grade and clinical stage.

OBJECTIVE: To investigate p53 and c-jun oncoproteins and proliferating cell nuclear antigen (PCNA) in transitional cell urinary bladder carcinomas (TCCs) and to determine their relationships to tumour grade, stage and survival. MATERIALS AND METHODS: The expression of p53, c-jun and PCNA was studied using immunohistochemistry in formalin-fixed, paraffin-embedded tissues in a series of 110 TCCs. RESULTS: 58% of our cases were positive for p53 and 88% for c-jun. A statistically very significant correlation (p < 0.0001) was observed between p53 and c-jun (r = 0.781), p53 and PCNA (r = 0.772), c-jun and PCNA (r = 0.831) as well as between each of the two oncoproteins and the histological grade and clinical stage (p < 0.001). There was no correlation of either p53, PCNA or c-jun with clinical outcome in terms of patients survival. CONCLUSION: p53 and c-jun proteins' overexpression are strongly related to rapid tumour cell proliferation and hence with aggressive growth in urinary bladder TCC. PCNA score remains an important prognostic index in transitional cell carcinoma of the bladder.