Repository logo

Browsing by Author "Bobrowska, Justyna"

  • Thumbnail Image
    Item
    Characterization of cell surface structure and its relation to cytoskeleton elasticity in cancer cells
    (Institute of Nuclear Physics Polish Academy of Sciences, 2016) Bobrowska, Justyna; Lekka, Małgorzata; Silberring, Jerzy; Lankosz, Marek; Laidler, Piotr
    The alterations observed in tumour cells include the number of processes introducing abnormalities in cellular morphology, structure and growth profiles. Despite continuous efforts, the molecular mechanism of the metastasis is still not understood completely. That is the reason why there is an urgent need for the search of new scientific approaches in the cancer progression investigations. The development of various biochemical and biological methods increases the chance to detect cancer, however, in past decades, single cell biomechanics has gained large significance since certain diseases are known to manifest in altered biomechanical properties. Stiffness of single cells is one of the major properties that changes during cancerous progression. Studies have demonstrated that biomechanics can bring both data describing mechanisms underlying cancer progression and tools for its detection and diagnosis at the single cell level. However, one of the emerging directions is to correlate cellular biomechanics with biochemical and biophysical properties of single cells.The main objective of the presented thesis is to study how single cell deformability is linked with cellular surface properties, and how these changes correlate with cancer progression. Thus, the elasticity of melanoma cells was measured by means of atomic force microscopy (AFM). Measurements were carried out for three groups of cells encompassing cells originating from primary tumour sites i.e. from radial and vertical growth phases (RGP and VGP, respectively), and those derived from skin and lung metastasis. The results were compared with properties of melanocytes (cells from which melanoma originates). The surface properties were determined using time of flight secondary ions mass spectrometry(ToF SIMS). The use of ToF SIMS has required to develop a dedicated sample preparation protocol enablingmeasurements of biological material in the high vacuum environment. The final results show the correlation between single cell deformability and surface biochemical properties of melanoma cells. They confirm the hypothesis that cancer progression causes alterations in the morphological and mechanical properties of cancerous cells and these differences are connected with changes in the cellular surface composition.
  • Thumbnail Image
    Item
    Multimodal approach for ionizing radiation damage investigation
    (Institute of Nuclear Physics Polish Academy of Sciences, 2015) Kwiatek, Wojciech M.; Adamczyk, Jolanta; Bobrowska, Justyna; Borkowska, Anna; Lekka, Małgorzata; Lekki, Janusz; Lipiec, Ewelina; Miszczyk, Justyna; Pabijan, Joanna; Paluszkiewicz, Czesława; Panek, Agnieszka; Piergies, Natalia; Pogoda, Katarzyna; Prauzner-Bechcicki, Szymon; Quaroni, Luca; Roman, Maciej; Wiecheć, Anna; Wiltowska-Zuber, Joanna
    The new, leading edge laboratory has been established and put into routine operation. The laboratory is focused on multimodal studies of the cellular response to ionizing radiation and provides support and research facilities for the Bronowice Cyclotron Centre. Spectroscopic imaging of cells and tissues (examining in particular the internal structure of cells, the cytoskeleton organization, cells’ mechanical and biochemical properties), as well as research at the molecular level, has been applied in research for the purpose of searching new strategies of prevention and therapies of cancer and other pathologies and in fundamental research in the field of structural and vibrational analysis of condensed matter. The multimodal approach, illustrating the full laboratory potential, has been applied to radiation effect studies of transitional cells – human bladder carcinoma cells (T24 cell line). Cells were irradiated with three X-ray radiation doses: 1 Gy, 3 Gy, and 10 Gy and subsequently studied using all instruments of the new laboratory. The presented results demonstrated that the AFM elasticity measurements can be applied to quantitatively estimate alterations induced upon Xray irradiation at the single cell level. A combination of AFM and InfraRed Spectroscopy (NanoIR2 setup) was successful in characterization molecular changes occurring in the nuclear environment following cellular irradiation. Particularly useful information acquired was the observation of changes in distribution of macromolecules with a spatial resolution at the level of the single organelle. The observed changes correlate with radiation dose and thus may become a tool for studying the biological effects of radiation exposure. The application of Raman microspectroscopy for radiation-induced damage investigations provided detection of such spectral changes as strand breaks, base unstacking, and DNA conformational transformations. These studies give crucial information about the damage associated with irradiation and cellular response for radiation dose at single cell level. Moreover, Raman spectroscopy provides possibility to investigate structural changes present within the same sample. Complementary UV–VIS technique, on the basis of γ-H2AX test, delivered quantitative data of radiation damage, manifesting in presence of double strand breaks in DNA in 1st and 2nd day of culture (1 hour and 24 hours after irradiation).