Antonina Harłozińska

Expression of the Tyrosine Kinase Activity Growth Factor Receptors (EGFR, ERB B2, ERB B3) in Colorectal Adenocarcinomas and Adenomas

The overexpression of three growth factor receptors: epidermal growth factor receptor (EGFR), ERB B2 and ERB B3 was evaluated immunohistochemically in 77 malignant and 15 benign colorectal neoplasms considering clinicopathological variables (histological structure, grade of differentiation, tumor localization, clinical stage of the disease). The relationship between the coexpression of EGFR-related proteins in individual patients was also evaluated. EGFR expression was revealed in comparable percentages of colorectal adenoma and in adenocarcinoma cases (80% and 70%) while ERB B2 expression was detectable more frequently in adenoma than in adenocarcinoma cases (87% and 54%). The presence of ERB B3 was observed in a higher percentage of adenocarcinoma than adenoma cases (65% and 40%). There was no correlation between the expression of studied tyrosine kinase receptors and histological grade or Dukes’ clinical stage and localization (proximal or distal) of colorectal adenocarcinoma. The incidence of EGFR and ERB B2 expression was higher in tubulovillous (100% for both receptors) than in tubular adenomas (63% and 75%), while the ERB B3 receptor was revealed more frequently in tubular than in tubulovillous neoplasms (50% and 28%). These differences appeared to be statistically nonsignificant. The concomitant expression of two growth factor receptors was observed in a higher percentage of colorectal adenomas than adenocarcinomas, and the coexistence of three growth factors was revealed in comparable percentages in malignant and benign colorectal tumors. Our results support the promotional rather than direct transformational role for the EGFR supergene family in colorectal tumorigenesis. The frequently observed coexpression of more than one EGFR-related protein in colorectal neoplasms indicates the possible cooperation of these receptors in mitogenic signaling transduction, facilitating the development and maintenance of the malignant phenotype.

Comparative analysis of CA125, tissue polypeptide specific antigen, and soluble interleukin-2 receptor α levels in sera, cyst, and ascitic fluids from patients with ovarian carcinoma

The serum markers CA125, tissue polypeptide specific antigen (TPS), and soluble interleukin‐2 receptor alpha (sIL‐2Rα) concentrations were determined in sera, cyst, and ascitic fluids from patients with malignant and benign ovarian neoplasms.

Field effect of human colon carcinoma on normal mucosa : Relevance of carcinoembryonic antigen expression

Human colon cancer usually develops on a mucosa which has already undergone multiple steps of genetic change. These multiple steps create a field effect characterized by the presence of morphologically normal, but biologically altered epithelial cells. This aims of this study were to evaluate whether the expression of carcinoembryonic antigen (CEA) can act as a phenotypic marker of the field effect, and to map its topography in relation to the presence of colorectal adenocarcinoma. The expression of CEA was tested by immunohistochemistry on morphologically normal mucosa at 4 increasing distances from 14 autologous cases of colorectal adenocarcinoma. CEA expression in the normal mucosa was compared to the tumor. The results show that in the mucosa adjacent to the edge of the autologous tumor, CEA is expressed to the same level as displayed in the carcinoma; there is a decrease in CEA expression in normal mucosa located at 1 cm or more from the edge of carcinoma. Mucosa sampled at 5 and 10 cm from the tumor expresses CEA at the same low level as in mucosa of control subjects with no colorectal neoplasm. In conclusion, this study demonstrates a gradient of CEA expression in the peritumoral area, supporting the concept of field effect, and maps its extent. These data are relevant to the biology of human colorectal cancer, and more practically, to the optimal location of surgical resection.

Epidermal Growth Factor Receptor and c-erbB-2 Oncoproteins in Tissue and Tumor Effusion Cells of Histopathologically Different Ovarian Neoplasms

The overexpression of two growth factor receptors – epidermal growth factor receptor (EGFR) and c-erbB-2 – was evaluated immunohistochemically in malignant and benign ovarian neoplasms, considering the stage of the disease and histology of tumors. The comparison of EGFR and c-erbB-2 reactivity in tissue sections and respective cyst and/or ascitic fluid cells was also performed. c-erbB-2 expression was detected in 44.4% of ovarian carcinomas, and in benign neoplasms there was no evidence of its staining, while EGFR reactivity was found both in malignant (58.7%) and benign (50%) tumors. Significant heterogeneity of staining was observed, however, the relationship between EGFR and c-erbB-2 expression in tissue sections and cyst and/or ascitic fluid cells in individual patients was evident. The expression of both growth factor receptors was not correlated with histopathological subtypes of ovarian neoplasms. The c-erbB-2 oncoprotein was detected more frequently in III/IV than in I/II stages according to the criteria of the International Federation of Gynecology and Obstetrics (FIGO) and the EGFR expression was independent of the clinical advancement of the disease. The coexpression of c-erbB-2 and EGFR was shown in 32% of ovarian carcinomas, and it dominated in cases with FIGO stages III/IV. Our results indicate that the increase of the EGFR expression appears to be associated with early stages of ovarian tumorigenesis, and the enhancement of c-erbB-2 reactivity may cooperate with EGFR activation in the development and progression of ovarian carcinomas.

nm23 expression in tissue sections and tumor effusion cells of ovarian neoplasms

The genetic changes involved in the metastatic process of ovarian epithelial cancer remain undetermined. The expression of nm23, a putative metastasis‐suppressor gene product, was assessed immunohistochemically in malignant and benign ovarian neoplasms, considering histology of tumors and clinical advancement of disease. Comparison of nm23 protein content in tissue sections and respective cyst and/or ascitic fluid cells was also performed. Significant heterogeneity of nm23 immuno‐staining was observed, and no correlation with histological subtype of ovarian carcinoma was found. Expression of nm23 was higher in carcinomas compared with benign tumors. A significant trend to have a higher nm23 reactivity in ascitic fluid cells vs. primary tumors was observed. Our results indicate that the increase of nm23 reactivity is activated in the early stages of the disease and that the progression of ovarian carcinoma is accompanied by overexpression of nm23 protein. Our observations did not confirm the postulated role of nm23 as a suppressor gene in ovarian cancer.

Expression and Mutation of p53 in Tumor Effusion Cells of Patients with Ovarian Carcinoma: Response to Cisplatin-Based Chemotherapy

p53 alterations are considered as one of the most important factors responsible for drug resistance in ovarian carcinomas, although the relationship between p53 gene status and response to cisplatin-based chemotherapy in ovarian cancer patients remains unclear. The aim of the study was to evaluate the relationship between p53 protein accumulation, p53 gene mutation and response to cisplatin-based chemotherapy in patients with ovarian carcinoma considering conventional clinicopathological parameters. Tissue sections and corresponding cyst and/or ascitic fluid cells from 79 patients with epithelial ovarian cancer were analyzed immunohistochemically for p53 expression. The PCR-SSCP analysis was performed in 25 cases and the results were compared with immunohistochemical data. It was demonstrated that p53 expression reaching approximately 50% of positive cells in immunostaining was usually associated with PCR-amplified exons showing abnormal migration and suspected for mutation. p53 gene changes were not correlated with histological structure, grade of differentiation or residual tumor after cytoreductive surgery, despite being detected more frequently in III/IV than in II FIGO stages and in patients with residual disease above 2 cm. A significant correlation between p53 accumulation and p53 gene alteration and poor response to cisplatin-based chemotherapy was shown. The overall survival time of patients decreased with an increase in p53 protein expression. A strong p53 expression especially accompanied by p53 changes detectable by PCR-SSCP analysis appears to be a good indicator of the resistance to cisplatin-based chemotherapy. The association between strong p53 overexpression and shorter overall survival time was also revealed.

Relation between ovarian carcinoma-associated antigens in tumor tissue and detached cyst fluid cells of patients with ovarian neoplasms.

Aims The expression and potential diagnostic value of ovarian carcinoma-associated antigens were estimated in different types of epithelial ovarian neoplasms. The comparison of antigenic expression was performed on solid tumor tissues and loose cyst fluid cells in individual cases of malignant and benign ovarian neoplasms. Methods All studies were performed using monoclonal antibodies (mAbs) against ovarian carcinoma-associated antigens (OC125, OV-TL3, OV632, 10B, 8C) by 3-step peroxidase-antiperoxidase test. Results All ovarian carcinoma-associated antigens were detected in most serous and endometrioid carcinomas. In mucinous carcinomas as well as in benign ovarian neoplasms these antigens were present only in some cases. Significant inter- and intratumoral immunological heterogeneity was evident; however, the antigens detectable in tissue sections were also found in detached cyst fluid cells. Conclusions Our results show that mAb show OV-TL3 is the best marker for endometrioid carcinomas and confirmed that mAbs OV632, OC125 and OV-TL3 could be good complementary markers for differentiating malignant and benign lesions in the ovary. The percentage content of all ovarian carcinoma-associated antigens in solid tumors and respective cyst fluid cells was comparable.

CYTOMORPHOLOGIC CHARACTERIZATION OF CELL SUBSETS ISOLATED BY DENSITY GRADIENT CENTRIFUGATION FROM TUMOR EFFUSIONS OF OVARIAN ENDOMETRIOID CARCINOMA

Aims Cytomorphologic characterization of tumor cell subsets, according to the stage of pathologic differentiation, and comparison of cellular composition in tumor cyst and ascitic fluids were carried out on individual patients with ovarian endometrioid carcinoma. Methods A density gradient centrifugation technique was applied to fractionate the cells from tumor effusions. Results The enrichment of cell forms representing individual stages of pathologic differentiation by gradient centrifugation facilitated their cytomorphologic characterization. According to cytomorphologic features, 5 discrete cell subpopulations were identified and catalogued. The cellular composition of tumor cyst and ascitic fluids in individual patients was similar, but the number of fractions and percentage of cell subsets differed. Conclusions The estimation of precise cytomorphologic criteria for cell forms in tumor effusions facilitated the cytologic diagnosis of ovarian endometrioid carcinoma. The possibility to concentrate poorly differentiated, frankly malignant cell subsets in low densities could significantly improve the diagnosis of tumor effusions.

Carcinoembryonic Antigen Isotypes in Tissue Sections and Loose Cyst Fluid Cells of Ovarian Neoplasms

The aim of this study was to establish whether different subsets of ovarian neoplasms express a restricted isotype of carcinoembryonic antigen (CEA) which can be detected in solid tumors and detached cells. Sixty-one cases of mucinous, serous, endometrioid, and Krukenberg tumors were studied by immunohistochemistry using two monoclonal antibodies (MAbs), commercial anti-CEA and D14 with a higher specificity for colorectal adenocarcinomas. The results with both antibodies showed a considerable degree of heterogeneity between cases of nonserous tumors, with a more restrictive pattern observed with the D14 MAb. The proportion of immunostained cells was comparable in tumors and fluids.

Reactivity of polyclonal and two monoclonal antibodies with cell subsets isolated from cystic fluids of ovarian serous neoplasms

The distribution of antigenic determinants on a cellular level in serous ovarian neoplasms was evaluated using polyclonal and two monoclonal antibodies (OC 125 and 10B). The expression of antigens was estimated by an immunofluorescence test on each cell fraction isolated by density centrifugation from cystic fluids of individual malignant and benign ovarian tumors, taking into account the density and cytomorphologic features of cell subpopulations. It was found that the studied antibodies recognized different antigenic determinants. Significant immunologic heterogeneity of cells among and within individual tumors was shown. Our studies show the value of isolated cell subpopulations for comparing the reactivity of different antibodies and estimating their immunodiagnostic potency.