BREAST TISSUE: BIOLOGICAL MECHANISMS OF DYSPLASIA AND GENETIC CRITERIA FOR DIAGNOSIS

layers; 2nd - moderate hyperplasia, when epithelial cells form more than four layers, often with concomitant closure of the hollow space; 3-4 - hyperplasia, if cell proliferation is observed in groups of hyperplastic cells with the typical appearance of overlapped and fuzzy nucleus distributions; atypical ductal hyperplasia morphologically mimics ductal preinvasive cancer; 5th - in


UKRAINE
The breast precancerous diseases have recently become widespread. Diagnostic studies of the breast today are focused on the detection of focal neoplasms. At the same time, it is proved that for 56% of the examined atypical breast hyperplasia occurs without nodule formation, therefore the frequency of cytological diagnosis errors in patients with benign breast tumors reaches 7%, and puncture uninformativeness -18.6% [1,2].
At the growing incidence of proliferative forms of mastopathy in recent decades and the young age of those suffering from these processes, it remains important to find mechanisms for early diagnosis and the latest criteria for assessing precancerous diseases. However, traditional invasive methods for assessing clinical and instrumental parameters and morphological changes are not informative enough. Therefore, the search for molecular genetic predictors of precancerous diseases is becoming increasingly important, which determines the relevance of this problem.
The aim of the work was to develop criteria for the diagnosis of proliferative benign breast dysplasia on the basis of immunohistochemical and molecular genetic studies in order to substantiate the indications for surgical treatment.
The general name of benign changes in the breast, which differ significantly in anatomical and clinical features, the risk of malignancy, makes us consider this disease as a precancerous disease -benign breast dysplasia (BBD).
There are several degrees of breast tissue proliferation with an increase in its atypia and the transition to cancer: 1st -the norm, where epithelial cells are arranged in three -four layers; 2nd -moderate hyperplasia, when epithelial cells form more than four layers, often with concomitant closure of the hollow space; 3-4 -hyperplasia, if cell proliferation is observed in groups of hyperplastic cells with the typical appearance of overlapped and fuzzy nucleus distributions; atypical ductal hyperplasia morphologically mimics ductal preinvasive cancer; 5th -in situ.
High and low levels of epithelial atypia in patients with BBD are determined. According to epidemiological studies, a high degree of atypia is associated with the development of breast cancer.
We studied breast tissue and blood. For histological examination, the material was taken during surgery and placed in a container with formalin. Blood from the peripheral vein was taken before or after surgery according to standard methods and stored at -20° C.
The presence of estradiol was studied by analyzing the combination of competitive enzyme-linked immunosorbent assay and fluorescent determination of reaction products, blood plasma analysis was performed. Conducted instrumental and laboratory studies. The morphological and immunohistochemistry features of the removed tissues, as well as genetic differences of patients were studied.
Ultrasound and mammography were performed during the BBD diagnosis [3]. Analysis of ultrasound and mammography data for each tumor was performed separately. Ultrasound examination was performed using an ultrasonic diagnostic system "Toshiba" Nemio XG SSA-580A, made in Japan (multifrequency linear sensor with a frequency of 6-12 MHz). Mammography was performed with a Hologic Lorad M-IV with a Kodak Direct View Classic CR digitizer (USA).
134 breast neoplasms received from 84 operated patients were studied. In 72 (53.7%) cases, samples from one organ were subjected to morphological examination, and in 62 (46.3%) neoplasms removed with bilateral breast lesions were analyzed. The mean time of preclinical observation of the subjects was (2.78 ± 0.161) years. In this case, the average size of the pathological tumor was equal to (22.29 ± 1.02) mm.
The results of the distribution of morphological samples of BBD by age show: most of the surgical drugs 94 (70.2%) removed for BBD belonged to young patients (up to 40 years); 40 (29.8%) morphological samples belonged to patients over 40 years of age. A significant number of morphological samples are due to multiple lesions of the breast. The study was dominated by BBD samples with proliferative activity of 3-4 degrees and metaplasia in some areas or samples with a tendency to atypical changes in 86 (64.2%) subjects. Neoplasms with unexpressed proliferative activity of 1-2 degrees were a minority -48 (35.8%) studies.
Venous blood from the studied patients was collected under sterile conditions in monovets with a volume of 2.7 ml with potassium salt of ethylenediaminetetraacetic acid (11.7 mm) -(Sarstedt, Germany), frozen and stored at -20 o C.
In patients with a mastopathy proliferative form, breast tissue obtained suboperatively at the border of healthy tissue -the affected area was studied. Histological examination was performed to evaluate the morphological changes of neoplasms in operated patients with BBD after surgery. All pathohistological specimens were divided into samples with mild (stage 1-2) and severe (stage 3-4) proliferation. For 65 (48.5%) drugs the fibroepithelial type of proliferation prevailed. On Fig.1 shows the proliferating stromal cells that compress the ducts with the formation of slit-like structures.
The stromal component is particularly pronounced in tumors of young women, often with significant proliferation of connective tissue cells along with myxomatosis, as well as hyalinosis and calcification. The epithelial component shows varying degrees of proliferation. There are foci of apocrine and squamous cell metaplasia. Myoepithelial type of proliferation prevailed in 15 (11.2%) drugs. The stromal component is particularly pronounced in young women tumors. The epithelial component shows varying degrees of proliferation. There are foci of apocrine and squamous cell metaplasia. Myoepithelial type of proliferation prevailed in 15 (11.2%) drugs.
The drug has tortuous, elongated or obliterated glands and tubules bounded by myoepithelial cells in the sclerosed stroma. Apocrine epithelial metaplasia is not excluded. At the massive defeat the so-called nodular or tumoral adenosis is formed. Epithelial partial and epithelial duct proliferation in the study occurred in 25 (18.7%) and 29 (21.6%) operated, respectively.
Despite the typical histological structure of tumors of different types of proliferation at the BBD, different levels of EsRα expression were found in all variants of the studied samples [4]. It was found that the frequency of epithelial lobular proliferation was 1.5 times higher for receptor-positive EsRα expression than for receptor-negative; epithelial duct proliferation -1.6 times, respectively, fibroepithelial proliferation was 1.2 times more frequent. Among the studied BBD samples neoplasms, the type of proliferation did not depend on the receptor status for EsRα expression (Fig.2).
Different proliferative variants of BBD with almost the same histological structure differ significantly in the expression of EsRα and the proliferation degree. This may reflect the peculiarities of hormonal status, as well as be a manifestation of breast cancer and an indicator of susceptibility to them. Prospects for the study of PvuIIpolymorphism in patients with precancerous pathology of the breast