Expression of ezrin and moesin in primary breast carcinoma and matched lymph node metastases.

Ezrin, radixin, moesin (ERM) are important membrane-cytoskeletal crosslinkers and are suggested to play important role in cancer progression and metastasis. Even though ERM proteins were generally considered to be functionally redundant and the most studied was ezrin, recent studies highlight their distinct roles in metastatic process. Little information is available regarding the role of individual ERM proteins and their phosphorylated forms in human breast cancer. Our study is the first to examine expression of ezrin, moesin and their phosphorylated forms in primary breast tumors and matched lymph node metastases (LNMs) and their correlation with clinicopathological variables. A total of 88 primary breast cancer, 91 LNMs, 54 intraductal carcinoma and 26 normal adjacent breast tissue samples from tissue microarrays were studied. Expression was determined by immunohistochemistry, the intensity and number of positive cells was scored. Statistical analysis of protein expression and patients' age, tumor grade and hormonal status was performed. No statistical significant difference was found in ezrin, moesin, p-ezrinTyr353 and pan-p-ezrinThr567/radixinThr564/moesinThr558 expression between primary tumors and LNMs. Even though it was not significant, moesin expression varied between primary tumors, intraductal carcinoma, normal breast adjacent tissue and LNMs. A significant positive correlation between moesin and tumor grade has been proven. Even though primary tumors and matched LNMs did not show different expression patterns, moesin correlated significantly with higher tumor grade. Its positivity in intraductal carcinoma and normal breast tissue adjacent to cancer might indicate its role in tumor intiation/progression.

[Functional morphology of recently discovered telocytes inside the female reproductive system]. / Funkcná morfológia novo objavených telocytov v zenskom pohlavnom systéme.

Discovery of telocytes has become an important and key challenge in past few years. These cells are interstitial cells extending very long cytoplasmic processes named telopodes, by which they create functional networks in the interstitium of different organs. Telocytes are considered to be connective tissue elements that create contacts among each other, but they also function as intercellular structures, functionally connected with cells of the immune system, neurons and smooth muscle cells. Telocytes can be found also in the different parts of female reproductive system with functions and purpose, which is summarized in our overview. Telocytes regulate for example peristaltic movements in fallopian tubes. The decrease of their number (due to inflammatory disease or endometriosis) causes impairment in transport through fallopian tubes which may result in sterility or tubal gravidity. In uterus they regulate contraction of myometrial smooth muscle (blood expulsion in menstrual phase, childbirth) as well as they contribute in immunological care during embryo implantation. Telocytes probably control also the involution of uterus after delivery. Their function in vagina has not been yet clearly defined; they probably take part in slow muscle contraction movement during sexual intercourse. In mammary glands some scientists suppose their function in control of cell proliferation and apoptosis, that is why, they may play a role in carcinogenesis. In placenta they probably monitor and regulate flow of blood in vessels of chorionic villi and they may be responsible also for etiopathogenesis of pre-eclampsy. All these mentioned functions of telocytes are only in the level of hypothesis and have been published recently. New research and studies will try to answer the questions whether telocytes play a key role in these processes. Our review we completed with some original microphotographs of telocytes in different organs of female reproductive system.

Prediction of additional lymph node involvement in breast cancer patients with positive sentinel lymph nodes.

Axillary lymph node dissection (ALND) has traditionally been the principal method for evaluating axillary lymph node status in breast cancer patients. In the past decades sentinel lymph nodes biopsy after lymphatic mapping has been used to stage the disease. The majority of sentinel lymph nodes (SLN) positive patients do not have additional metastases in non-sentinel nodes (non-SLN) after additional ALND. These patients are exposed to the morbidity of ALND without any benefit from additional axillary clearence. In the present study we would like to asses the criteria for selecting those patients, who have high risk for non-SLN metastases in the axilla in cases of positive SLN. In this retrospective analysis, clinical and pathologic data from 163 patients who underwent SLN biopsy followed by ALND were collected. Following clinical and pathological characteristics were analyzed to predict the likehood of non-SLN metastases: age, staging, histologic type and grading of the tumors, hormonal receptor status, HER-2 receptor status and Ki-67 protein, angioinvasion, metastases in SLN and non-SLN. Relative frequencies of individual characteristics between sample groups were statistically tested by Chi-square test at significance level p=0.5, when sample sizes in groups were small (≤5) by Fisher´s exact test. Metastasis in SLN were present in 67 (41%) of patients, 48 patients (29,4%) had metastasis also in non-SLN. The ratio between non-SLN positive / non-SLN negative lymph nodes in patients with positive SLN increases with the stage of the disease, the difference between values for the pT1c and pT2 stadium was statistically significant (p = 0.0296). The same applies to grading, but the differences were not significant (p>0.05). We could not find significant differences for angioinvasion of the tumor, probably for small number of patients with angioinvasion (p>0.05).Only the stage of the tumor was shown to be significant in predicting the metastasis in non-SLN in our group of breast cancer patients with positive SLN Nearly 80% of the patients of 70 years and older displayed no benefit from axillary staging, because of negative SLN as well as non-SLN, although thanks to the small sample size this was not a statistically significant result. Furthermore, current recommendations for axillary staging in breast cancer patients are discussed.

COX-2, p16 and Ki67 expression in DCIS, microinvasive and early invasive breast carcinoma with extensive intraductal component.

BACKGROUND: Recent studies have showed a significant association between the combination of COX-2, p16 and Ki67 overexpression and incidence of subsequent invasive carcinoma in a subgroup of treated ductal carcinoma in situ (DCIS) and the indicated prognostic value of COX-2, p16 and Ki67 in early breast cancer. Based on the continual model of carcinogenesis and the mentioned results, we hypothesize, that if COX-2, p16 and Ki67 expression is prognostic for DCIS future behaviour, the expression level of the markers correlates also with different stages of breast carcinomas such as DCIS, microinvasive cancer and early invasive cancer with an extensive intraductal compound. The aim of this study was to compare the expression of COX-2, p16 and Ki67 in different stages of breast carcinoma such as pure DCIS, microinvasive cancer (T1mic) and invasive ductal carcinoma with an extensive intraductal component (IDC with EIC). The expression was assessed only in in situ component of the three subgroups (DCIS, T1mic, EIC) in order to show a possible correlation of COX-2, p16 and Ki67 with different stages of carcinogenesis. METHODS: We carried out a retrospective study using immunohistochemical staining to evaluate the expression of the markers COX-2, p16 and Ki67 in in situ lesions within three subgroups of tumors with the rising extant of invasive compound: in pure DCIS, microinvasive carcinoma (T1mic) and invasive carcinoma with extensive in situ component (IDC with EIC). Additionally, we performed a correlation analysis between the tumor subgroups and patients history data (age, parity, age of menarche, family and personal cancer history, breast feeding lengths, contraception intake, chest irradiation) as well as some of the tumor characteristics (tumor grade, multicentricity, necrosis). RESULTS: Distribution of p16 expression differed significantly among the three diagnoses. P16 score 1 was highest in the DCIS group whereas the lowest proportion was in IDC and p16 overexpression (score 2, 3) maintained this tendency (overexpression proportion in DCIS < T1mic < IDC), though this was not significant. The frequency of COX-2 and p16 overexpression (phenotype COX-2+p16+) was higher in EIC within invasive carcinoma in comparison to DCIS and T1mic and was rising gradually with the severity of the diagnosis (proportion in DCIS < T1mic < IDC). CONCLUSION: This is the first published study ever assessing the expression of COX-2, p16 and Ki67 markers in different breast tumors containing DCIS compound. Our results showed an increasing expression pattern of COX-2 and p16 with the rising severity of the diagnosis (expression was measured exclusively in in situ lesions within tumors containing different extant of invasiveness). The same relationship was showed for p16 marker alone. These data support different expression pattern of COX-2 and p16 markers in combination and p16 marker alone in "in situ lesions" according to the stage of carcinogenesis. This fact might be useful in the evaluation of further behaviour of early breast tumors (Tab. 3, Fig. 8, Ref. 29).

Local recurrence rate in patients with DCIS.

UNLABELLED: The aim of this observational retrospective study was to evaluate the local recurrence rate of ductal carcinoma in situ of the breast (DCIS) and/or invasive breast cancer in patients with DCIS or microinvasive carcinoma of the breast after breast conserving or radical surgery. Secondary aim of the study was comprehensive assessment of the whole management of DCIS and its comparison with European guidelines. METHODS: The study was performed in a group of 41 women with DCIS or microinvasive cancer, who underwent surgical treatment (breast conserving or radical modified mastectomy) at the IInd Department of Gynaecology and Obstetrics, University Hospital Bratislava (UNB), during the period 2001-2009. Documentation and pathological examination data from paraffin embedded tissue sections were used as data source. We sent out questionnaires regarding data about additional postoperative treatment and course of the disease up to year 2010 with focus on recurrence or tumour de novo incidence. RESULTS: Breast conserving surgery was performed in 28 cases - 68 %, modified radical mastectomy in 13 cases. All cases of mastectomy were due to multicentricity and/or extensive tumour >4 cm. Additional surgery due to unsatisfactory marginal status was performed in 8 patients (3.28 %). Additional treatment such as radiotherapy and/or hormonal therapy received 19 patients. Van Nuyss Prognostic Index was reported in 17 patients on the basis of histopathological data. 27 patients completed and returned questionnaire. No DCIS recurrence nor infiltrating cancer or tumour de novo was reported in this group. CONCLUSION: We consider surgical management as adequate. Further material analysis is needed (Tab. 5, Fig. 1, Ref. 28).

Semi-quantitative RT-PCR assessment of molecular markers in breast large-core needle biopsies.

Large-core needle biopsies are frequently used for the preoperative evaluation of the breast lesions. In addition to initial diagnostic information, they can show the status of molecular markers with predictive and prognostic value and further contribute to an optimal selection of treatment strategy. So far, the potential use of large-core needle biopsies in assessment of marker profile of the breast lesions was studied using the immunostaining approaches. In this work, we sought to determine whether analysis of the large-core needle biopsy by semi-quantitative reverse-transcription polymerase chain reaction reveals molecular data that correspond with the marker status of the subsequently removed tumor. Five molecular markers including ER, PR, c-erbB-2/HER-2/neu, c-erbB-3/HER-3, and CD44 were assessed on mRNA isolated from 23 large-core needle biopsies and corresponding surgical breast cancer specimens using beta2- microglobulin as an internal standard. Significant or highly significant correlation between core biopsies and excised tumors was observed for each marker when the mRNA expression status was scored as positive or negative, with the concordance of the data ranging from 73.9% to 86%. Using the dichotomous scoring, majority of the biopsies (75%) displayed molecular profile that was either identical to the profile of the related tumor specimen, or with the difference in one marker. However, no significant correlation was found when the levels of the markers were expressed as continuous variables, possibly due to intratumoral cell heterogeneity. These results suggest potential usefulness and reliability of semi-quantitative RT PCR in the evaluation of large-core needle biopsies with regard to marker positivity or negativity. On the other hand, the marker-related data expressed as continuous variables cannot be accurately assessed on the large- core needle specimens using this approach, indicating the need for methodological improvements.

The normal female and the male breast epithelium does not express prostate-specific antigen. Preliminary immunohistochemical observations of autopsy breast tissues.

In the normal female and male breast epithelial structures any prostate-specific antigen (PSA) immunohistochemical positivity was observed. Variable PSA expression, which often borders the positivity, was observed in membranes of adipocytes of fat tissue and in the endothelium of small vessels in a female and a male breast. Based on these initial observations, tissue of the normal breast, male or female, can not be considered to be the principal source of PSA.

[Phospholipids in the human myometrium in various stages of contraction before and during labor]. / Fosfolipidy humánneho myometria pri rôznych stavoch kontrakcnej cinnosti pred termínom a v termíne pôrodu.

A detailed analysis of the myometrium showed that the physical state of phospholipids--fluidity--depends on the ratio of their individual components and changes in relation to the state of contractile activity. The results indicate that before term of labor changes in the fluidity of myometrial phospholipids as well as increased availability of arachidonic acid for prostaglandin synthesis can induce preterm onset of contractile activity. Possibilities of affecting these mechanisms, which belong to the many potential factors inducing preterm labor, are discussed.

[Lipid composition of the myometrium during labor]. / Lipidické zlozky myometria pocas pôrodnej cinnosti.

Lipids represent one of the basic components of each cellular and subcellular membrane of the myometrium and their fluidity has a strong influence upon membrane function. Human myometrium was obtained at cesarean sections. Lipids were separated by one-dimensional thin layer chromatography. The chromatoplates were determined on the densitometer Shimadzu CS 930. Lipid profile of the myometrium was studied before the 37th week of pregnancy, at term without contractile activity, further during at term labor with normal contractile activity and at failure of myometrial contractility. Analysis of the obtained data showed changes in lipid fluidity, namely a decrease before the 37th week of pregnancy and at failure of myometrial contractions during at term labor. The decrease of fluidity was caused by a higher content of total cholesterol and a lower content of total phospholipids in the myometrium.