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The comparison of properties of cell lines resistant to ellipticine, doxorubicin, and cisplatin
Černá, Tereza ; Poljaková, Jitka (advisor) ; Eckschlager, Tomáš (referee)
7 Abstract Neuroblastoma is the most common extracranial solid tumor of childhood. Despite advances in cancer diagnosis and therapy, the treatment of some forms of neuroblastoma is still complicated. One of the major complications of the chemotherapy is a developed drug resistance. This master thesis deals with the effect of cytostatics on protein and gene expression of selected proteins, which may contribute to chemoresistance of the human neuroblastoma cell line UKF-NB-4. The sensitive line UKF-NB-4 and the resistant line UKF-NB-4CDDP , UKF-NB-4DOXO and UKF-NB-4ELLI were exposed to cisplatin, doxorubicin, ellipticine for 24, 48 and 72 hours. The Western blot analysis showed that cytostatic agents cisplatin, doxorubicin or ellipticine added to the sensitive neuroblastoma cell line UKF-NB-4 in amounts which are added to resistant neuroblastoma cell lines in order to maintain resistance induced expression of p53 and reduced expression of retinoblastoma protein pRb after 72 hours of cultivation. Differences in the expression of RAS protein, cytochrome P450 1A1, 3A4 and cytochrome b5 has not been shown. Changes in the expression of the studied proteins in resistant lines UKF-NB-4CDDP , UKF-NB-4DOXO and UKF-NB-4ELLI cultured with and without cytostatic agents were not detected by the Western blot analysis....
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Effects of natural compounds on viability of tumor cell lines
Boleslavská, Barbora ; Poljaková, Jitka (advisor) ; Bořek Dohalská, Lucie (referee)
Cancer is considered to be one of the most serious issues of medicine nowadays. Moreover, its incidence is still rising. Despite the huge progress in modern treatment methods, cancer therapy is still limited by many difficulties. This work focuses on the natural substances such as epigallocatechin gallate, caffeine, Cannabis sativa ethanol extract, Origanum acutidens water extract, Mentha piperita water extract and its effects on the human neuroblastoma cell line UKF-NB-4. The first part of the bachelor thesis deals with determining the viability of human neuroblastoma cell line UKF-NB-4 exposed to tested substances as well as it studies the effects of those substances on the cell cycle and caspase activity. Finally, it was tested, whether those substances are able to induce apoptosis in neuroblastoma cell lines. Tests were undertaken on the MUSETM cell analyzer and on the flow cytometer. The second part of the bachelor thesis focuses on the expression of protein p53 and retinoblastoma protein in neuroblastoma cell lines exposed to tested substances. Detection was carried out by Western blot analysis. Epigallocatechin gallate exhibited the most significant effect on human neuroblastoma cell line. It lowers the expression of retinoblastoma protein as well as it arrests cells between G0/G1 and S...
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The expression of macrophage migration inhibitory factor in neuroblastoma cell lines
Polatová, Daniela ; Poljaková, Jitka (advisor) ; Groh, Tomáš (referee)
A macrophage migration inhibitory factor (MIF) is a cytokine which participates in immune responses induced by outer stimuli such as lipopolysaccharides of bacteria or glucocorticoids. It is produced mainly by macrophages. It activates other macrophages and T-lymphocytes and allows anti-inflammatory cytokines to be released. It contributes to angiogenesis - endogenously produced MIF is crucial for the proliferation of endothelial cells. The formation of new blood vessels stimulated by MIF was also described in tumors and it leads to tumor growth. The natural receptor for MIF is an integral membrane protein CD74, which could be presented on the cell surface. If it is presented on the surface of T-lymphocytes, it could be modified by chondroitin- sulphate and interact with a protein CD44. This modification and interaction is important for an effective activation of T-cells. MIF then binds to the complex CD74-CD44. The presence of CD74 in human neuroblastoma cell lines UKF-NB-3 and UKF-NB-4 was studied in this bachelor thesis. Using electrophoresis and Western blot CD74 was detected in the UKF-NB-4 neuroblastoma cell line but it wasn't detected in the UKF-NB-3 neuroblastoma cell line. Nevertheless, the presence of this receptor in neuroblastoma cells indicates, that even neuroblastomas could be...
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The expression of macrophage migration inhibitory factor in neuroblastoma cell lines
Polatová, Daniela ; Poljaková, Jitka (advisor) ; Groh, Tomáš (referee)
A macrophage migration inhibitory factor (MIF) is a cytokine which participates in immune responses induced by outer stimuli such as lipopolysaccharides of bacteria or glucocorticoids. It is produced mainly by macrophages. It activates other macrophages and T-lymphocytes and allows anti-inflammatory cytokines to be released. It contributes to angiogenesis - endogenously produced MIF is crucial for the proliferation of endothelial cells. The formation of new blood vessels stimulated by MIF was also described in tumors and it leads to tumor growth. The natural receptor for MIF is an integral membrane protein CD74, which could be presented on the cell surface. If it is presented on the surface of T-lymphocytes, it could be modified by chondroitin- sulphate and interact with a protein CD44. This modification and interaction is important for an effective activation of T-cells. MIF then binds to the complex CD74-CD44. The presence of CD74 in human neuroblastoma cell lines UKF-NB-3 and UKF-NB-4 was studied in this bachelor thesis. Using electrophoresis and Western blot CD74 was detected in the UKF-NB-4 neuroblastoma cell line but it wasn't detected in the UKF-NB-3 neuroblastoma cell line. Nevertheless, the presence of this receptor in neuroblastoma cells indicates, that even neuroblastomas could be...
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Effects of natural compounds on viability of tumor cell lines
Boleslavská, Barbora ; Poljaková, Jitka (advisor) ; Bořek Dohalská, Lucie (referee)
Cancer is considered to be one of the most serious issues of medicine nowadays. Moreover, its incidence is still rising. Despite the huge progress in modern treatment methods, cancer therapy is still limited by many difficulties. This work focuses on the natural substances such as epigallocatechin gallate, caffeine, Cannabis sativa ethanol extract, Origanum acutidens water extract, Mentha piperita water extract and its effects on the human neuroblastoma cell line UKF-NB-4. The first part of the bachelor thesis deals with determining the viability of human neuroblastoma cell line UKF-NB-4 exposed to tested substances as well as it studies the effects of those substances on the cell cycle and caspase activity. Finally, it was tested, whether those substances are able to induce apoptosis in neuroblastoma cell lines. Tests were undertaken on the MUSETM cell analyzer and on the flow cytometer. The second part of the bachelor thesis focuses on the expression of protein p53 and retinoblastoma protein in neuroblastoma cell lines exposed to tested substances. Detection was carried out by Western blot analysis. Epigallocatechin gallate exhibited the most significant effect on human neuroblastoma cell line. It lowers the expression of retinoblastoma protein as well as it arrests cells between G0/G1 and S...
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The comparison of properties of cell lines resistant to ellipticine, doxorubicin, and cisplatin
Černá, Tereza ; Poljaková, Jitka (advisor) ; Eckschlager, Tomáš (referee)
7 Abstract Neuroblastoma is the most common extracranial solid tumor of childhood. Despite advances in cancer diagnosis and therapy, the treatment of some forms of neuroblastoma is still complicated. One of the major complications of the chemotherapy is a developed drug resistance. This master thesis deals with the effect of cytostatics on protein and gene expression of selected proteins, which may contribute to chemoresistance of the human neuroblastoma cell line UKF-NB-4. The sensitive line UKF-NB-4 and the resistant line UKF-NB-4CDDP , UKF-NB-4DOXO and UKF-NB-4ELLI were exposed to cisplatin, doxorubicin, ellipticine for 24, 48 and 72 hours. The Western blot analysis showed that cytostatic agents cisplatin, doxorubicin or ellipticine added to the sensitive neuroblastoma cell line UKF-NB-4 in amounts which are added to resistant neuroblastoma cell lines in order to maintain resistance induced expression of p53 and reduced expression of retinoblastoma protein pRb after 72 hours of cultivation. Differences in the expression of RAS protein, cytochrome P450 1A1, 3A4 and cytochrome b5 has not been shown. Changes in the expression of the studied proteins in resistant lines UKF-NB-4CDDP , UKF-NB-4DOXO and UKF-NB-4ELLI cultured with and without cytostatic agents were not detected by the Western blot analysis....
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