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název v anglickém jazyce není uveden
Krásná, Luboslava ; Veselý, Pavel (advisor) ; Křepela, Evžen (referee) ; Červinka, Miroslav (referee)
An in vitro cultivation technique has been developed that allows to reproducibly obtain and propagate a heterogenous population of epithelial cells from samples of normal mammary glands, breast tumours and subcutaneous metastases of breast cancer. The epithelial origin of cultured cells was proved by their cytokeratin profile, and EMA and ESA protein expression. The technique uses a feeder layer of irradiated NIH 3T3 cells to support clonal growth of even single isolated cells. Cultures were successfully established from 96% of surgical tumour biopsy specimens (69 out of 72, volume of sample about 1 cm3) and 67% of true-cut biopsy specimens (28 out of 42, volume 0.01 cm3). Two and more in vitro passages were obtained with 74% (53 out of 72) of surgical specimens and 31% (13 out of 42) of truecut biopsy specimens. This is the first report of successful in vitro propagation of cells obtained from breast cancer via true-cut biopsy. In cell populations analyzed for the growth properties, the culture lifetime, related to the number of colony-forming cells, varied for cells from benign tumours between 22 and 40 cell populations and for malignant tumours between 21 and 51. The population doubling time varied betwen 18 and 62 hours for benign cultures and 10 and 127 hours for malignant cultures, with average 32...
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Molecular mechanisms of apoptosis induction in tumor cells
Koc, Michal ; Kovář, Jan (advisor) ; Červinka, Miroslav (referee) ; Haškovec, Cedrick (referee)
V. Závěr Tato práce se zaměřila na studium procesů uplatňujících se v indukci apoptosy různými faktory. V našem případě jsme indukovali buněčnou smrt u nádorových buněk deprivací železa, klinicky používanými taxany (paclitaxel, docetaxel) a nově syntetisovanými fotosensitivními látkami. Z presentovaných publikací jsme získali tyto závěry. (1) Studie týkající se indukce apoptosy deprivací železa prokázaly účast odlišných signálních drah vedoucí k indukci apoptosy u myších a lidských nádorových buněk. V obou případech byla použita B lymfomová buněčná linie citlivá k deprivaci železa. U myších buněk jsme popsali významnou část apoptotické signální dráhy u 38C13 buněk sensitivních k indukci apoptosy deprivací železa. Deprivace železa u těchto buněk vede k translokaci proapoptotického proteinu Bax z cytosolu na mitochondrie, která je následována zhroucením mitochondriálního membránového potenciálu v důsledku porušení integrity vnější mitochondriální membrány, uvolněním cytochromu c z mitochondrií, aktivací kaspasy 9 a následnou aktivací kaspasy 3. U lidských buněk jsme taktéž zaznamenali indukci apoptosy deprivací železa, avšak charakterem mechanismu odlišnou od apoptosy popsané u myších 38C13 buněk. V případě lidských Raji buněk sensitivních k indukci apoptosy deprivací železa docházelo pouze k aktivaci kaspasy...
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Molecular mechanisms of apoptosis induction in tumor cells
Koc, Michal ; Kovář, Jan (advisor) ; Červinka, Miroslav (referee) ; Haškovec, Cedrick (referee)
V. Závěr Tato práce se zaměřila na studium procesů uplatňujících se v indukci apoptosy různými faktory. V našem případě jsme indukovali buněčnou smrt u nádorových buněk deprivací železa, klinicky používanými taxany (paclitaxel, docetaxel) a nově syntetisovanými fotosensitivními látkami. Z presentovaných publikací jsme získali tyto závěry. (1) Studie týkající se indukce apoptosy deprivací železa prokázaly účast odlišných signálních drah vedoucí k indukci apoptosy u myších a lidských nádorových buněk. V obou případech byla použita B lymfomová buněčná linie citlivá k deprivaci železa. U myších buněk jsme popsali významnou část apoptotické signální dráhy u 38C13 buněk sensitivních k indukci apoptosy deprivací železa. Deprivace železa u těchto buněk vede k translokaci proapoptotického proteinu Bax z cytosolu na mitochondrie, která je následována zhroucením mitochondriálního membránového potenciálu v důsledku porušení integrity vnější mitochondriální membrány, uvolněním cytochromu c z mitochondrií, aktivací kaspasy 9 a následnou aktivací kaspasy 3. U lidských buněk jsme taktéž zaznamenali indukci apoptosy deprivací železa, avšak charakterem mechanismu odlišnou od apoptosy popsané u myších 38C13 buněk. V případě lidských Raji buněk sensitivních k indukci apoptosy deprivací železa docházelo pouze k aktivaci kaspasy...
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Study of new prognostic markers in patients with chronic lymphocytic leukemia
Vrbacký, Filip ; Červinka, Miroslav (advisor) ; Pospíšilová, Šárka (referee) ; Procházka, Vlastimil (referee)
3 Summary Title: Study of new prognostic markers in patients with chronic lymphocytic leukemia Chronic lymphocytic leukemia (CLL) is the most common leukemic disorder of adults in Western hemisphere. It is characterized by clonal proliferation and accumulation of morphologically mature lymphocytes in bone marrow, peripheral blood and lymphatic tissues. Clinical course of CLL is extremely heterogeneous with some patients living for decades without need of therapy while others succumbing to the disease within several years. Thus, there has been great interest in identifying prognostic markers that could be used to distinguish patients with an aggressive form of CLL, because they might benefit from early intervention. The process of angiogenesis has been shown to be crucial for growth and metastasizing ability of solid tumors. Angiogenesis in "liquid" tumors was supposed to be less important, nevertheless numerous recent studies have shown enhanced angiogenesis in many hematological malignancies including CLL. Elevated levels of circulating angiogenic cytokines and increased expression of genes encoding angiogenic factors have been reported in recent years in patients with chronic lymphocytic leukemia (CLL) but data regarding their prognostic and predictive significance are still limited. Therefore, in the...
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Chemosenzitivity testing of human ovarian cancer cells and its in vitro model, human ovarian cancer cell line A2780
Caltová, Kateřina ; Červinka, Miroslav (advisor) ; Vaňásek, Jaroslav (referee) ; Hofmanová, Jiřina (referee)
Ovarian cancer is the fifth most common cancer and the most frequent cause of the death among gynaecological cancers in population of Czech women. The latest data from the National oncological register of Czech Republic show the incidence 20,64 and mortality 12,58 in 100 000 women (2010). A serious problem is represented by a late diagnosis of cancer, when the disease is diagnosed in the late stadium and finally this causes high mortality from this kind of cancer. Most complicated factors include high heterogenity of the disease, frequent apperance of resistance to cytostatics and low individualization of chemotherapy. In cooperation with a Clinic of gynaecology and obstetrics, Faculty hospital in Hradec Kralove, we have tested solid tumors and ascites samples of patients with ovarian cancer diagnosis to predict answers to chemotherapy within in vitro conditions. A panel of six cytostatics (cisplatin, paclitaxel, carboplatin, gemcitabine, topotecan, etoposide), which are also used in primary chemotherapy of ovarian cancer, has been tested. We have also used a model system, a human ovarian cancer cell line A2780, co compare the reactivity to tested cytostatics. Our results show the highest sensitivity of cells isolated from clinical samples of solid tumors and ascitic fluid to topotecan and...
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The mechanism of cell death induction in human malignant melanoma in vitro
Rudolf, Kamil ; Červinka, Miroslav (advisor) ; Mlejnek, Petr (referee) ; Janisch, Roman (referee)
Melanoma is a complex malignant disease characterized by increasing incidence and mortality. Melanoma biology is complicated and comprises many aberrant genes and signaling pathways. Treatment of melanoma relies on surgery, the role of radiation therapy and chemotherapy are limited. In advanced disseminating and metastasizing melanoma chemotherapy largely fails and that is why new approaches are being sought including identification of deregulated molecules or processes which could be used as targets. Some of these targets center on frequently aberrant cell death regulating mechanisms. Some traditional cytostatic drugs such as inhibitors of topoisomerases although useful in other types tumors so far proved useless in treatment of melanoma. Nevertheless, expression of topoisomerases in melanoma is often high, thereby enabling at least theoretically their use as targets for chemotherapeutical intervention. Aim of this study was to assess the cytoxicity and proapoptotic effect of campthotecin (topoisomerase I inhibitor), etoposide (topoisomerase IIα inhibitor) and their combination in selected human melanoma cell lines. We found that campthotecin may induce apoptosis by a combination of mechanisms including p53 as well as p73 and kaspase-2. Etoposide-induced cell death in Bowes melanoma cells included...
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název v anglickém jazyce není uveden
Krásná, Luboslava ; Veselý, Pavel (advisor) ; Křepela, Evžen (referee) ; Červinka, Miroslav (referee)
An in vitro cultivation technique has been developed that allows to reproducibly obtain and propagate a heterogenous population of epithelial cells from samples of normal mammary glands, breast tumours and subcutaneous metastases of breast cancer. The epithelial origin of cultured cells was proved by their cytokeratin profile, and EMA and ESA protein expression. The technique uses a feeder layer of irradiated NIH 3T3 cells to support clonal growth of even single isolated cells. Cultures were successfully established from 96% of surgical tumour biopsy specimens (69 out of 72, volume of sample about 1 cm3) and 67% of true-cut biopsy specimens (28 out of 42, volume 0.01 cm3). Two and more in vitro passages were obtained with 74% (53 out of 72) of surgical specimens and 31% (13 out of 42) of truecut biopsy specimens. This is the first report of successful in vitro propagation of cells obtained from breast cancer via true-cut biopsy. In cell populations analyzed for the growth properties, the culture lifetime, related to the number of colony-forming cells, varied for cells from benign tumours between 22 and 40 cell populations and for malignant tumours between 21 and 51. The population doubling time varied betwen 18 and 62 hours for benign cultures and 10 and 127 hours for malignant cultures, with average 32...
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