Detection of cell proliferation index is widely used in experimental and clinical research. Earlier it was shown that nuclear Ki-67 protein expression is strictly related to cell proliferation. It was revealed during all active phases of the cell cycle in mammals but was absent in G0 phase, so Ki-67 presence in cell nuclei reflects a potential growth fraction of whole cell population. The main area of Ki-67 antibody application is in immunocytochemical and immunohistochemical analyses. The aim of our work was to generate mouse monoclonal antibodies for Ki-67 antigen detection in mammalian tissues and in cultured cells. His-tagged fragment of Ki-67 expressed in bacteria was used as an antigen. Antibody-producing hybridoma cells were generated by standard procedure by fusing SP2/0 myeloma cells with splenocytes of immunized mice. Monoclonal antibodies were analyzed using paraffin-embedded human melanoma tissue samples and breast cancer cell line MCF-7. It was shown that generated anti-Ki-67 antibodies revealed proliferating cells in MCF-7 culture and after heat-induced epitope retrieval on paraffin sections of human melanoma tissue. In summary, generated antibodies could be useful for detection of proliferating cells in immunohistochemical and immunofluorescence studies of mammalian cells and tissues.

A scratch test is one of the most popular methods of classical cell migration assay in a monolayer culture. At the same time, the scratch assay has some disadvantages that can be easily corrected. Aim. Optimization the existent scratch assay on the base of detection of scratch wound surface area and the length of the field of observation which is more objective and less time consuming. Methods. Scratch assay. Results. The modification of scratch assay enables to perform measurement more accurately and rapidly. This approach is more simple and eliminates the main disadvantages of the classical method. Conclusions. The procedure of scratch wound width measurement calculated on the base of detection of cell free area and the length of the field of observation is more effective than the classical wound healing assay. It will be useful for the estimation of cell migration dynamics in monolayer culture for a wide range of live cell based experiments.

The main functional compartments of molecular chaperone Hsp60 are mitochondria and cytoplasm. Up to 30 % of Hsp60 are located in cytoplasm of cardiomyocytes. The interaction between molecular chaperone Hsp60 and proapoptotic Bax protein in the cytoplasmic fraction from normal human heart tissue has been revealed by co-immunoprecipitation in contrast to myocardium affected by dilated cardiomyopathy, where this interaction has not been observe

In this study two cDNA expressing libraries generated from thyroid papillar carcinomas were screened using SEREX approach. Thirty positive cDNA clones representing seventeen different genes were identified from both libraries. It is important to note, that three of them were isolated previously by other laboratories in SEREX screens of various types of human cancer. These include transcription factor NZF, a-catenin and ВАС RPU-a protein with unknown function. Moreover, we identified a whole panel of novel potential tumor-associated antigens, which would be further investigated. We are particularly interested in more detailed analysis of cathepsin H and transducer of ErbB2 (TOB2), which are differentially expressed in various types of human cancer. We will analyse the frequency of autoantibodies against identified antigens in sera of patients with various malignancies and healthy donors by heterologous screening. It is expected that among the clones isolated in this study, there might be novel cancer-associated markers.

Migration capacity is an important feature of tumor cells. There are several approaches to analyze the dynamics of cancer cell migration in vitro. The model of initiation of cell migration from 3D multicellular spheroids onto growth surface is one of the closest to the in vivo conditions. Aim. Optimization of the model to study tumor cell mobility for several days. Methods. 2D and 3D MCF-7 cell culture, immunofluorescence analysis and image analysis using the Fiji computer software. Results. Unification of spheroid size allowed avoiding a significant data deviation. The obtained spheroids spread completely for three days. The highest migration ratio was observed on the second day. The proliferation level was similar during each day of the three-day experiment; it did not exceed 3 %. The validity of the model was tested after migration inhibition by a mTOR signaling inhibitor rapamycin. Additionally, this model was successfully applied to immunofluorescence study of p85S6K1 subcellular localization in moving MCF-7 cells. Conclusions. Double filtration of multicellular spheroids allowed unification of their size; this promotes an adequate interpretation of the migration assay. This model allows to study tumor cell migration dynamics and can be further used for development of anticancer drugs.

Для пошуку нових пухлинних антигенів раку щитовидної залози людини використано метод SEREX. Ідентифіковано 16 різних антигенів шляхом скринування аутологічними сироватками двох експресуючих кДНК бібліотек, виділених з тканин папілярних карцином раку щитовидної залози. Здійснено алогенний скринінг одержаних антигенів сироватками здорових донорів, а також пацієнтів, хворих на рак щитовидної і молочної залоз та товстого кишечника. Вісім антигенів прореагували виключно з аутологічними сироватками, два (KY-Thy-25 та KY-Thy-40)-лише з сироватками пацієнтів, хворих на рак щитовидної і молочної залоз (10 та 2,4 % відповідно), а решта антигенів прореагувала як із сироватками здорових донорів, так и хворих на рак. KY-Thy-17 (білок, який містить мотив «цинковий палець» типу С2Н2), KY-Thy-28 (переносник ERBB2) та KY-Thy-29 (білок, асоційований з солідними пухлинами) виявили високу імунореактивність із сироватками пацієнтів, хворих на рак щитовидної та молочної залоз і раком товстого кишечника, у порівнянні з сироватками здорових донорів. Згадані антигени можуть бути перспективними для розробки на їхній основі нових діагностичних та імунотерапевтичних підходів при ракових захворюваннях. Вступ. Рання діагностика та імунотерапія пухлин базується на пошуку пухлинних антигенів, здатних викликати гуморальну та клітинну імунну від повідь у онкологічних хворих. Пухлиноасоційовані антигени, идентифіковані на сьогодні, можна роз поділити на п'ять групп: 1) ембріоспецифічні анти гени (CT-cancer-testis), які експресуються у різних типах пухлин і відсутні в нормальних тканинах, за винятком сім'яників; 2) антигени диференціації меланоцитів, що експресуються в меланомах і нор мальних меланоцитах; 3) антигени, які виникли © Р.

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