[Determination of receptor and HER2 status of breast cancer using tissue matrix technology. Validation of the method].

We studied ability of TMA technique as alternative to analysis of whole sections (WS) for breast carcinomas. The study included 229 patients with breast carcinoma T1-2 N0 M0 treated within 2000-2009 years. After whole slide scanning were constructed TMA blocks using 2mm punches and TMA master device (3DHistech, Budapest). TMA results of immunohistochemistry of HER2 were compared retrospectively with the results of WS. General distribution of HER2 status for WS was: negative (0+1) 85,5%, unequal (2+) 6,07%, positive (3+) - 8.41%, for TMA cores negative (0+1) 82,6%, unequal (2+) 7,04%, positive (3+) - 10,33%. When we compared each cases for HER2 there was moderate agreement with negative (score 0) and positive(score 3+) case around 70% and poor agreement in HER2 (1+2+ category) - only 50% 25% including both false negative and false positive estimations. We thought such kind disagreement in individual group was due to using different primary antibodies (all not FDA-approved) in previous years compared with PATHWAY® rmAb 4B5 (Ventana) antibody which was used for TMA slides. For breast cancer TMAs could be reasonable alternative to WS. Especially for Her2 testing, should be used only FDA approved systems and this technique allow reduce time, reagent and finally cost of such type of analysis.

[Obesity and characteristics of endometrial cancer: are there any changes over several decades?].

It is shown that endometrial cancer features (including expression of the PTEN and HER-2/neu proteins) are connected dissimilarly with body mass index and with the belonging of the patients to the groups with standard, SO and metabolically healthy obesity, MHO. In the course of the last half-century an increases are discovered in the height and weight of the females with endometrial cancer that moves in the opposite direction with a reduction of the share of MHO cases among obese patients. This conclusion should be taken into account when one considers the means for contemporary prevention of both obesity and cancer of uterine body.

[METHODOLOGIC PROBLEMS OF SENTINEL LYMPH NODE BIOPSY IN PATIENTS WITH BREAST CANCER].

The study included data on 168 patients with breast cancer, surgical treatment of whom was supplemented by axillary dissection (133 patients or 79.2%) or biopsy of sentinel lymph nodes (35 patients or 20.8%). The examination included ultrasound, planar scintigraphy of the breast and zones of regional lymph drainage. In 122 patients with primary breast cancer stage cT1-2N0M0 retrospective analysis of radionuclide imaging sentinel lymph node was performed. In 89 patients the introduction of colloidal radiopharmaceutical was carried out using a particle diameter of not more than 80-100 nm, in 33 patients study was conducted after administration of radiocolloid with a particle diameter of 200 to 1000 nm. Based on the data obtained by scintigraphy and ultrasonography of zones of regional lymph drainage there were offered two diagnostics models: the first, in which the presence of metastatic axillary lymph nodes was established when there were changes according to at least one of the diagnostic methods--scintigraphy or ultrasound; the second, in which the defeat of lymph nodes was determined only in the case of simultaneous detection of ultrasound and scintigraphic evidence of axillary lymph nodes. Sensitivity, specificity, and overall accuracy of the combination of ultrasound and planar scintigraphy axillary lymph nodes using the first model accounted for 82.7%, 67.7% and 74.4%, respectively. In the second model, the specificity was 94.6%, sensitivity--56%. Rapid transport of radiopharmaceuticals from the injection site, a high gradient of radiopharmaceuticals accumulation in sentinel lymph nodes, effective their visualization, approaching to 100%, were undoubted advantages of radiocolloids having a particle diameter up to 100 nm.

[The role of scintimammography with 99mTs-technetril to predict the pathologic status of regional lymph nodes in patients with locally advanced breast cancer after neoadjuvant chemotherapy].

The purpose of this study was to establish the role of scintimammography with 99mTs-technetril for predicting pathological status of regional lymph nodes after neoadjuvant chemotherapy. In 123 primary patients with locally advanced breast cancer stage cT1-4N0-3M0 there was performed scintimammography: before treatment and after 2-3 cycles of neoadjuvant chemotherapy 63 patients; after 2-3 cycles of neoadjuvant chemotherapy and before surgery--in 5 patients; after 4-6 cycles of neoadjuvant chemotherapy--55 women. All patients were divided into 2 groups: the first included 68 patients whom scintimammography was performed before and after 2-3 cycles of neoadjuvant chemotherapy; the second--60 women whom scintimammography was performed before treatment and before surgery. In patients of the first group positive results were obtained in 16 (23.5%), negative--26 (38.3%), false positive--16 (23.5%) and false negative--10 (14.7%) cases. Sensitivity, specificity, overall accuracy, positive predictive value and negative results were 61.5%, 61.9%, 61.7%, 50% and 72.2%, respectively. During the scintimammography after 4-6 cycles of neoadjuvant chemotherapy positive results were observed in 17 (28.3%) negative--18 (30%), false positive--19 (31.7%) and false negative--6 (10%) of patients. Thus, the sensitivity, specificity, overall accuracy, positive predictive value and negative results amounted to 73.9%, 48.6%, 58.3%, 47.2%, 75% respectively.

[Immunohistochemical prognostic factors of primary refractory Hodgkin's lymphoma].

In order to identify immunohistochemical markers associated with primary refractory Hodgkin's lymphoma, 20 patients with primary refractory form at stage IIAB in complete remission underwent histological examination and immunohistochemical determination of Bcl-6, Bcl-2, c-kit (CD117), CD15, CD30, P-53, Ki-67. In the group with primary refractory Hodgkin's lymphoma at stage IIAB Bcl-6 expression was found in 2 patients (10%), Bcl-2 in 14 patients (70%), c-kit (CD117) in 16 patients (80%), CD15 in 9 patients (45%). The expression of CD30 and P-53 was observed in all patients in this group (100%). The expression of Ki-67 ranged from 20% to 100%, 80-100% in 16 patients (80%). As a result, multivariate analysis revealed no immunohistochemical markers of primary refractory Hodgkin's lymphoma. In univariate and multivariate analyzes in a group of primary refractory Hodgkin's lymphoma a high level P-53 expression (80-100%) was significantly associated with decreased overall survival. A 5-year overall survival in patients with Hodgkin's lymphoma with the expression level of P-53 < 80% was 78%, with the expression level of P-53 80-100%-22%. A 5-year overall survival of Hodgkin's lymphoma primary patients in complete remission significantly exceeded the rates of patients with primary refractory Hodgkin's lymphoma--100% vs. 52%.

[Effectiveness of mammary scintigraphy for determining the results of neoadjuvant polychemotherapy in breast cancer].

Analysis of mammoscintigraphy is presented to assess the effectiveness of neoadjuvant polychemotherapy for locally advanced breast cancer. In 90% of patients there is defined conformity of scintigraphy data on the effectiveness of performed polychemotherapy to the results of histologic examination of the postoperative material. The sensitivity of scintigraphy in assessing the effectiveness of neoadjuvant polychemoterapy was 83.3%, specificity--87.9%, accuracy--86.3%, predictive value of positive and negative results-78.9% and 90.1%, respectively. Mammoscintigraphy is an effective method to control the response of locally advanced breast cancer to neoadjuvant polychemotherapy.

[Family diabetes and its consequences in cancer patients].

Preliminary data are confirmed on the more rare prevalence of family history of diabetes mellitus (DM) in cancer patients, mainly females, with diabetes in comparison with diabetics without cancer pathology. Familial diabetes does not worsen additionally tumor characteristics against the same in patients with non-familial diabetes. More than that, familial diabetes in diabetics with breast cancer goes together with lesser size of tumor and demonstrates an inclination to the rarer distant metastases in breast and endometrial cancer patients. The signs of systemic DNA damage (evaluated, in particular, on the basis of 8-OH-dG serum levels) are pronounced in postmenopausal diabetic cancer patients with familial diabetes in lesser degree than in non-familial variant of DM. In toto, this allows to consider family history of DM in patients with type-2 diabetes as a particular factor of tumor growth containment, which mechanisms and causes, warrant further studies.

[Markers of effectiveness of preoperative taxane-based chemotherapy for locally advanced breast cancer].

The absolute sensitivity signs of breast cancer to the drug have not yet been developed. Data from clinical trials on the study of experimental laboratory predictive markers of chemosensitivity: TOP2alpha (topoisomerase 2-alpha), beta-tubulin (subunit of dimeric protein tubulin), and BRCA1 (breast cancer 1) are contradictory and not numerous. Analysis of the results by the end of the clinical trial will allow examining the correlation between the effectiveness of preoperative taxane-chemotherapy and the level of experimental and standard molecular markets that is important for development of algorithm of treatment tactics for patients with locally advanced breast cancer.

[Metformin does not suppress the aromatase expression in breast cancer tissue of patients with concurrent type 2 diabetes].

Although there is data suggesting the in vitro inhibition of aromatase in cell lines by antidiabetic biguanide metformin (MF), there is no data on the intratumoral breast cancer (BC) aromatase expression in patients already receiving therapy for type II diabetes. Paraffinized tumor samples obtained from 57 BC pts aged 48-77 yrs, >80% of pts had stage T1-2N0-3M0 BC. Thirteen of the pts didn't have diabetes, 44 pts were previously diagnosed type II diabetes and reseaved the following therapy for at least 1 year: diet only (n=14), sulphonylurea (SU, n=14), metformin (MF, n=9) or MF with SU (n=7). Tumor samples were deparaffinized in xylene and treated with the monoclonal aromatase antibody 677. The rate and intensity of tissue staining was then analyzed by semi-quantitative method using conventional scores. Negative controls were processed with 0.01 M PBS instead of the specific antibody. For positive control paraffin-embedded human placenta samples were used. By conventional scores method the following values were obtained: 1.31 (pts without diabetes), 1.47 (all diabetic patients), 2.22 (MF), 1.50 (SU), 1.29 (MF+SU), 1.81 (MF and MF+SU), 1.07 (diet). Allred scores for progesterone receptor (PR) were the highest in the samples from pts treated with MF or MF+SU and the lowest in the samples obtained from SU-treated pts. Thus, in contrast to previous findings suggesting the suppressive effect of MF on aromatase in vitro, no such trend was discovered for aromatase expression in tumor samples from diabetic patients treated with MF. Although the investigated patients population is still small, this data combined with clinical data (higher PR levels) may suggest the better responses to hormonal therapy in MF-treated diabetic patients.

[Effectiveness of aromatase inhibitors in comparison with metformin for neoadjuvant treatment in patients with endometrial cancer].

We compared the efficacy of endometrial cancer neoadjuvant treatment in 38 patients receiving nonsteroid (letrozol, anastrozol) or steroid (ekzemestan) aromatase inhibitors and 12 patients receiving metformin. The changes in glucose metabolism were revealed in 26.3% of patients treated with aromatase inhibitors and 16.7% of patients treated with metformin. However, comparison of endometrial thickness (M-echo signal) data, postoperative data on favorable differentiation grade changes rate and proliferative activity (Ki-67 expression) revealed the superiority of 2-4 weeks aromatase inhibitors course in comparison with 2-9 (average 5.3 +/- 0.7) weeks metformin treatment.

[Effect of previous diabetes therapy on tumor receptor phenotype in breast cancer: comparison of metformin and sulphonylurea derivatives].

According to some existing data, unlike sulphonylurea (SU) and insulin derivatives, treatment with biguanide metformin, for reasons still unknown, may diminish breast cancer (BC) morbidity in diabetic females. For its part, diabetes is known to worsen survival of BC patients although there is no evidence of a pathway by which antidiabetic therapy might influence the key prognostic feature of BC tissue--the tumor receptor phenotype. Combination of BC and diabetes (n=90) was studied. SU drugs were received for at least 12 months by 22 patients, biguanide metformin alone or in conjunction with SU by 15, insulin by 5, and dietary treatment alone--by 48 pts. Percentage of estrogen receptor-positive tumors did not vary significantly from group to group. However, progesterone receptor-positive (PR+) tumors in metformin-treated patients were revealed more often than in those receiving SU alone (p = 0.43) or with insulin (p = 0.041), respectively. Hence, previous treatment with metformin is expected lead to higher incidence of PR+ tumors which in turn may stimulate efficiency of hormonal therapy only in relevant group of diabetic BC patients.

[A rare case of small-cell diffuse invasive breast lobular carcinoma].

The paper describes a rare case of invasive lobular carcinoma with the diffuse growth type simulating malignant non-Hodgkin lymphoma. The immunohistochemical study of the tumor established a marked expression of pancytokeratin, GSDFP-15, receptors to estrogen and progesterone, CD 7, Bcl-2, expression CD 3, CD 5, CD 10, CD 20, CD 23, CD 30, ALK, Bcl-6, inactive expression of E-cadherin. The ability of invasive lobular carcinoma to simulate malignant non-Hodgkin lymphoma determines the diagnostic value and significance of an immunohistochemical study to confirm the epithelial nature of the tumor.

[Criteria for evaluating the effectiveness of aromatase inhibitors in the neoadjuvant treatment of patients with endometrial carcinoma].

Clinical and experimental effects of neoadjuvant treatment of endometrial cancer patients with non-steroidal aromatase inhibitors: letrozole (femara, n=10, 2.5 mg/day, 14 days), anastrozole (arimidex, n=15,1 mg/day, 28 days) and exemestane (aromazine, n=13, 25 mg/day, 14 days) were compared. Administration of anastrozole was mostly frequently followed by pain relief in the lower abdomen and/or decreased rates of uterine discharge. Endometrial wall thickness (M-echo signal) decreased significantly in 60% of patients receiving anastrozole, exemestane - 58.3% and letrozole - 40%. Substantial drop in intratumoral aromatase and blood estradiol levels occurred more frequently after anastrozole and letrozole while progesterone receptor levels in tumor were markedly lower after exemestane administration. Assay of blood LH (except letrozole), FSH and cholesterol appeared to be of less relevance. On the contrary, significance of assessment of marker Ki-67 expression, which, in the case of anastrozole, dropped in 6 out of 12 patients after a 28-day course, could hardly be underestimated.

[Progenotoxic shift in mammary adipose tissue (adipogenotoxicosis): association with clinical and biological characteristics of breast cancer].

The study is concerned with identification of a relationship between levels of production and accumulation of compounds capable of hormonal and progenotoxic effects in mammary fat, on the one hand, and characteristics of tumor tissue in breast cancer, on the other. Mammary fat was sampled at a distance of 1.5-2 cm from tumor edge (79 pts.). Case histories were used to provide data on clinical stage, size, grade and regional lymph node involvement. Levels were assayed of leptin, adiponectin, tumor necrosis factor (TNF-alpha), interleukin-6 (IL-6), nitric oxide (NO), thiobarbiturate-reactive products (TBRP) and DNA oxidative damage marker (8-OH-dG) from 4hr-incubates of fat tissue culture. Mammary fat aromatase was assayed by radiometrical means while macrophage-assisted fat infiltration (CD68) and estrogen-4-hydroxylase (CYP1B1) expression were evaluated immunohistochemically. Radio-competitive and immunohistochemical methods were used to assay estrogen (ER) and progesterone (PR) receptor levels in tumor and tumor-related expression of cytokeratins 5/6 ("basal") and 7/8 ("luminal" epithelium), respectively. As far as hormonal properties of mammary fat were concerned, there were direct correlations between aromatase concentration, on the one hand, and tumor stage and size, on the other, and adiponectin secretion and CK7 expression in tumor. Besides, an inverse correlation was found between mammary fat-mediated release of leptin and adiponectin, on the one hand, and stage and regional lymph node involvement, on the other. The following main relationships were identified by comparison of the clinico-biological characteristics of tumor and markers of proinflammatory/progenotoxic properties of mammary adipose tissue: tendency toward direct correlation with IL-6 and 8-OH-dG in fat (tumor progress stage); direct correlation with TNF-alpha secretion rate (malignancy grade); lymph node involvement--tendency toward direct correlation with NO generation; CK5 expression in tumor--tendency toward direct correlation with 8-OH-dG, TBRP and CD68 fat infiltration; CK7 expression in tumor--tendency toward inverse correlation with NO generation in adipose tissue; ER-negative phenotype of tumor--tendency toward higher generation of TBRP, NO and TNF/leptin in fat. Hence, shift toward predominance of proinflammatory/progenotoxic properties of mammary adipose tissue (adipogenotoxicosis) is associated with signs of less favorable course of tumor process in the mammary gland which calls for working out adequate measures.

[An immunocytochemical investigation of antigen expression by skin melanoma cells cultured for vaccine production].

There is a great variety of histological patterns of skin melanoma and, in particular, that of its metastatic patterns. Malignant melanocytes are capable of influencing tumor-associated antigen expression. As of now, several varieties of melanoma-associated antigens (MAA) have been identified: MART1/melan A, tyrosinase, MITF, gp100, members of MAGE family, S100, CD63 and CD146. Peptides isolated from such molecules can induce MHC-restricted response of cytotoxic T-lymphocytes. It has been shown that level and nature of specific antigen expression caused by melanocytes correlate with tumor stage and a relationship between survival and MAA expression on tumor cells identified. Morphological features, growth pattern and proliferation rate varied in melanoma cell cultures used in our study. Our experiments involved evaluation of changes in the properties of antigens HLA (class 1 and 2), tumor tissue samples and cells isolated from them, which were capable of stable proliferative activity during passages 5, 10, 15, 20, 25, 30 and 35, and assay for MAA content. Levels of the antigens were significantly lower following long-term culturing melanoma cell melanoma cells in vitro. At initial passages (1-5), antigen profile in most cultures was similar to that in tumor tissue samples. Later on each cell population showed greater antigen expression heterogeneity matched by increased number of cells going through mitotic cycle; their nuclei were stained with antibodies to Ki-67. No HLF A/B/C molecule expression took place during tumor cell culturing: stained cells--in 68.9% of cultures (passages 1-5) and 36.3% (passage 35). However, HLA DQ/DP/DR molecule identification showed an inverse relationship: 44.1% (passage 5) while virtually all the cell lines did synthesize those molecules after passage 35. Hence, MAA and MHC (class 1 and 2) antigens expression in tumor cell should be monitored when they are used for preparation of autologuos and allogenic vaccines. In case of allogenic vaccine production, cell lines capable of stable production of MAA should be selected.

[Biological features of breast cancer in patients under 35].

The study was concerned with biological features of inoperable breast cancer and their prognostic significance for patients under 35. That age was identified as a prognosticator for breast cancer: high-grade (G3) malignancies, vascular invasion and EIC+component were relatively more frequent in such patients. Also, those with HER2/neu hyperexpression had worst prognosis.

[Clinico-radiological assessment of chemotherapy for osteosarcoma received prior to surgery]. / Kliniko-rentgenologicheskaia otsenka éffektivnosti khimioterapii osteosarkom v predoperatsionnom periode.

A clinico-radiological evaluation of chemotherapy received by 20 patients with osteosarcoma at the Institute's Clinic (1999-2003) was conducted. Morhological confirmation of the findings on course dynamics agreed upon by clinicians and roentgenologists was obtained in 78%. X-ray examinations provided more information. The clinico-radiological evidence on tumor regression failed morphological test in one-third of cases.

[Phase I-II trial of gene therapy with tag-7 modified autologous tumor cells in patients with metastatic solid tumors]. / I-II faza klinicheskoi otsenki éffektivnosti genoterapii na osnove autologichnykh opukholevykh kletok, modifitsirovannykh genom TAG7 u bol'nykh s disseminirovannymi solidnymi opukholiami.

A prospective (phase I-II) trial was undertaken to study the efficacy and toxicity of gene therapy with tag 70-modified autologous tumor cells in 32 patients with metastatic renal cell carcinoma (RC) (5) and melanoma (MBL) (27) treated at the Institute's Clinic (2001-2003). Resected material was reduced to cell culture, which was transfected with tag 70 gene and devitalized by irradiation. Immune blotting was used for gene expression. Clinical and immunological effectiveness was evaluated in 22 patients (MBL--17 and RC--5) who received 1-6 injections (3 on the average). Full course of vaccination was given to 8 (MBL--6 and RC--2). No complete or partial response was reported while least regression (50%) was registered in a case of RC metastatic to the lung. According to CT and ultrasound evidence, stabilization was achieved in 5 (23.8%) (MBL--4 and RC--1). Relapse-free period was 6.5+/-3.5 months beginning from the start of treatment. The vaccine was well tolerated while DHT reaction was observed in 47.6% (10 out of 17) of primary immunized patients. A trend of increased content of T- and B-cells in peripheral blood and intensified functional activity was established.

[An improved procedure for autologous gene-modified cancer vaccine preparation for active specific immunotherapy of disseminated solid tumors]. / Usovershenstvovanie metoda prigotovleniia autologichnykh modifitsirovannykh protivoopukholevykh vaktsin dlia aktivnoi spetsificheskoi immunoterapii bol'nykh s disseminirovannymi solidnymi opukholiami.

Resected material was used from 108 patients with disseminated skin melanoma and 47 patients suffering metastasized renal tumors to test procedures of tumor cell culture preparation and to search the best parameters. Gene tag 7 transfer, liposome delivery and electroporation were employed to stimulate immunogenic tumor cells. The transfer results were evaluated by expression of beta-galactosidase and EGFP genes whose products were detected microscopically. Transfer efficiency was boosted by 30% due to selecting suitable parameters of tumor cell modification. Maximum effectiveness was attained by individualized choice of the parameters. Yet, undoubtedly, the best way of cell isolation was mechanical fragmentation of tumor. To speed up cell production, DMEM/F12 medium should be recommended. It should contain cattle embryonic serum (20%), conditioned medium of cultured fibroblasts of human embryonic lungs (20%), transferin, insulin and selenium (standard dose).