Marek Kulikowski

Circulating free DNA, p53 antibody and mutations of KRAS gene in endometrial cancer

This study was conducted to evaluate the significance of circulating free DNA (CFDNA), p53 antibody (p53‐Ab) and mutations of KRAS gene in the development of endometrial cancer (EC). A total of 109 patients with EC (87 patients with Type I and 22 patients with Type II) took part in this study. KRAS mutations and CFDNA were detected by means of the PCR‐RFLP and enriched by the PCR‐RFPL method. ELISA was used to analyze plasma p53‐Ab. Tissue expression of P53 protein was evaluated immunohistochemically (IHC). The frequency of KRAS mutations was especially high in Grade 2 of Type I EC. CFDNA was frequently detected in patients with early stage of Type II EC at a low level of grade. It is noteworthy that the p53‐Ab positive rate increased in the higher grade of Type I tumors. A significant difference in the number of cases with the p53‐Ab was found in the advanced stage of Type I tumors. The frequency of KRAS and p53‐Ab correlates with tumor stage only in the Type I EC. Plasma CFDNA and p53‐Ab offer a chance to develop a procedure for EC Type II diagnosis. The association between tumor cells related to CFDNA and p53‐Ab with Type II tumor suggests that it might potentially serve as a marker in predicting the prognosis and offers a possibility to individualize treatment regimen.

Mutations of the KRAS oncogene in endometrial hyperplasia and carcinoma.

The aim of this study was to examine the prevalence and clinicopathological significance of KRAS point mutation in endometrial hyperplasia and carcinoma. We analysed KRAS in 11 cases of complex atypical hyperplasia and in 49 endometrial carcinomas using polymerase chain reaction associated with restriction fragment length polymorphism (PCR-RFPL). Point mutations at codon 12 of KRAS oncogene were identified in 7 of 49 (14,3%) tumor specimens and in 2 of 11 (18,2%) hyperplasias. No correlation was found between KRAS gene mutation and age at onset, histology, grade of differentiation and clinical stage. We conclude that KRAS mutation is a relatively common event in endometrial carcinogenesis, but with no prognostic value.

Serum levels of VEGF and VEGF-C in patients with endometrial cancer

Endometrial cancer (EC) is the most common type of uterine cancer. A dualistic model of endometrial tumorigenesis serves as a useful way of categorizing these cancers in terms of both etiology and clinical behavior. There are two types of EC: type I and type II. Type I is so-called estrogen-dependent, and appears mostly in pre- and perimenopausal women, it is well differentiated and therefore has a better prognosis. Type II EC is estrogen-independent, diagnosed mostly in postmenopausal women, thin and fertile women, or in women with normal menstrual cycles. It is aggressive and has a worse prognosis than type I. The aim of this study was to evaluate the relationship between the pretreatment serum levels of VEGF and VEGF-C and the outcome of EC patients. A total of 98 patients treated between 1999 and 2003 were included in this study. Circulating VEGF and VEGF-C levels were determined using ELISA kits. VEGF levels among the 76 patients with type I, and the 22 patients with type II EC were significantly higher than those found in the healthy control subjects (p < 0.001). The differences in mean values of VEGF-C were highly significant in both types of tumor examined compared to the control (p < 0.001). The results demonstrate that serum VEGF concentration correlated significantly with advanced FIGO stage in type II EC (p < 0.001). The preoperative VEGF-C level correlated with advancing tumor stages in type I EC (p < 0.05). An elevated preoperative VEGF-C was an independent risk factor for disease-specific survival in patients with type II tumors. Thus, in type II EC patients with high preoperative levels of VEGF-C, pelvic and para-aortic lymphadenectomy should be performed. However, the value of longitudinal measurements of the markers used is yet to be determined.

Mutations in the KRAS gene in ovarian tumors

RAS genes are the most frequently mutated oncogenes detected in human cancer. In this study we analyzed the presence of mutations at codon 12 of the KRAS gene in 78 women with ovarian tumor, including 64 invasive ovarian cancers and 14 borderline ovarian tumors, using an RFLP-PCR technique and we evaluated whether such alterations were associated with the selected clinicopathological parameters of the patients. KRAS codon 12 gene mutations were found in 6,2% of ovarian cancer tissue and in 14,3% of the borderline ovarian tumor. KRAS mutations were found with a significantly higher frequency in mucinous and borderline tumors compared to serous tumors (p<0,01). Mutation frequency was correlated with the histological type of tumor, but not with stage, grade or patients age.

The prognostic significance of the immunohistochemical expression of P53 and BCL-2 in endometrial cancer.

The objective of this study was to verify the frequency of P53 and BCL-2 immunohistochemical expression in 98 patients with endometrial carcinoma, and to correlate it with clinical stage and patient survival. A significant difference was found regarding the frequency of P53 expression when comparing type I and II tumors (23.7% and 54.5%, respectively; p = 0.006). A positive correlation was observed between P53 immunoexpression and patient survival in type I and II tumors (p = 0.009 and p = 0.036, respectively). BCL-2 expression was significantly more frequent in early clinical stages in both types of endometrial cancer (p 〈 0.001 and 0.002) and correlated with a decrease in overall survival in type I endometrial cancer (p = 0.014). Thus, the prognostic value of these biomarkers in endometrial cancer needs to be further investigated.

Prognostic significance of smac/DIABLO in endometrioid endometrial cancer

Apoptosis may occur via a death receptor-dependent or independent (mitochondrial) pathway. The mitochondrial pathway is regulated by small molecules, such as smac/Diablo, which activates caspase cascades. This study examined smac/DIABLO expression in 76 patients with endometrioid endometrial cancers. Presence of smac/DIABLO was quantified by Western blot analysis using nonfixed fresh frozen tissues. Its appearance was found in 55 (72%) of examined tumors. Smac/DIABLO expression significantly correlated with tumor grade (p<0.001). Patients with positive smac/DIABLO tumors had a longer disease-specific survival when compared with those with negative tumors in the 10-year follow-up (p=0.043). The study demonstrated that negative smac/DIABLO expression was a poor prognostic sign.

Antioxidant status in women who have had a miscarriage

PURPOSE During normal pregnancy there is an oxido-reductive balance between action of pro-oxidative factors. The aim of this study was to evaluate the total antioxidant status (TAS) and glutathione peroxidase (GSH-Px) activity, and the content of selenium (Se), zinc (Zn), copper (Cu) and manganese (Mn) in women who have had a miscarriage. PATIENTS AND METHODS The study group consisted of 83 women who had had miscarriages. The control group included 35 women in the first trimester of pregnancy, and 35 pregnant women after childbirth. RESULTS TAS activity and Cu concentration in serum in women who experienced a miscarriage were significantly lower, but Mn level - higher, than in women in the first trimester of pregnancy. The content of Se, Cu and Mn in placental tissue in patients who have had a miscarriage was significantly higher, while Zn content was lower than in pregnant women at full-term delivery. CONCLUSIONS Our findings, provided in the research, enable us to claim that the total antioxidative status is significantly lower in women who have had a miscarriage. Low level of Zn but high of Mn in the examined biological material may be indicative to the incidence of miscarriage.

Nucleolar Organizer Regions in Differentiated Preneoplastic and Neoplastic Endometrial Lesions

The aim of the study was the quantitative assessment of nucleolar organizer regions (NORs) in the cells of preneoplastic and neoplastic endometrial lesions. Retrospective analysis included tissue material obtained from 87 patients with simple and complex hyperplasia with or without atypia and well-differentiated stage-I endometrial adenocarcinoma. Sections were made from routine paraffin blocks and stained with AgNO3. The mean number of AgNORs per cell was determined. The results obtained indicate that the evaluation of proliferative activity of cells expressed by AgNOR count can help in the distinction between atypical hyperplasia and well-differentiated adenocarcinoma, and thus can serve as a useful pathological criterion.