This bachelor's thesis deals with the development and basic characterization of cell lines resistant to nucleoside cytostatics, cultivated in vitro. The theoretical part of the work is devoted to general facts associated with the development of cancer and mechanisms of resistance to cytostatic drugs, focusing on nucleoside derivatives that are used in clinical practice for the treatment of leukaemia and solid tumours. The literature review also highlights the importance of in vitro models in drug development.
The beginning of the experimental part of the work is focused on the selection of resistant cells. Resistance was verified using the MTS cytotoxicity test, furthermore, the screening of new potential drugs is involved. The second part is dedicated to their basic characterization using standardized procedures for this laboratory. The expression of P-glycoprotein and MRP1 was analyzed by flow cytometry. The presence of proteins associated with the mTOR pathway was determined by western blotting.
Annotation in English
This bachelor's thesis deals with the development and basic characterization of cell lines resistant to nucleoside cytostatics, cultivated in vitro. The theoretical part of the work is devoted to general facts associated with the development of cancer and mechanisms of resistance to cytostatic drugs, focusing on nucleoside derivatives that are used in clinical practice for the treatment of leukaemia and solid tumours. The literature review also highlights the importance of in vitro models in drug development.
The beginning of the experimental part of the work is focused on the selection of resistant cells. Resistance was verified using the MTS cytotoxicity test, furthermore, the screening of new potential drugs is involved. The second part is dedicated to their basic characterization using standardized procedures for this laboratory. The expression of P-glycoprotein and MRP1 was analyzed by flow cytometry. The presence of proteins associated with the mTOR pathway was determined by western blotting.
Keywords
Drug resistance, nucleosides, oncology, in vitro models
Keywords in English
Drug resistance, nucleosides, oncology, in vitro models
Length of the covering note
63 s
Language
AN
Annotation
This bachelor's thesis deals with the development and basic characterization of cell lines resistant to nucleoside cytostatics, cultivated in vitro. The theoretical part of the work is devoted to general facts associated with the development of cancer and mechanisms of resistance to cytostatic drugs, focusing on nucleoside derivatives that are used in clinical practice for the treatment of leukaemia and solid tumours. The literature review also highlights the importance of in vitro models in drug development.
The beginning of the experimental part of the work is focused on the selection of resistant cells. Resistance was verified using the MTS cytotoxicity test, furthermore, the screening of new potential drugs is involved. The second part is dedicated to their basic characterization using standardized procedures for this laboratory. The expression of P-glycoprotein and MRP1 was analyzed by flow cytometry. The presence of proteins associated with the mTOR pathway was determined by western blotting.
Annotation in English
This bachelor's thesis deals with the development and basic characterization of cell lines resistant to nucleoside cytostatics, cultivated in vitro. The theoretical part of the work is devoted to general facts associated with the development of cancer and mechanisms of resistance to cytostatic drugs, focusing on nucleoside derivatives that are used in clinical practice for the treatment of leukaemia and solid tumours. The literature review also highlights the importance of in vitro models in drug development.
The beginning of the experimental part of the work is focused on the selection of resistant cells. Resistance was verified using the MTS cytotoxicity test, furthermore, the screening of new potential drugs is involved. The second part is dedicated to their basic characterization using standardized procedures for this laboratory. The expression of P-glycoprotein and MRP1 was analyzed by flow cytometry. The presence of proteins associated with the mTOR pathway was determined by western blotting.
Keywords
Drug resistance, nucleosides, oncology, in vitro models
Keywords in English
Drug resistance, nucleosides, oncology, in vitro models
Research Plan
Příprava rezistentních buněčných linií
Exprese proteinů spojených s mnohočetnou lékovou rezistencí
Screening nových potenciálních léčiv
Research Plan
Příprava rezistentních buněčných linií
Exprese proteinů spojených s mnohočetnou lékovou rezistencí
Screening nových potenciálních léčiv
Recommended resources
Noskova V, Dzubak P, Kuzmina G, Ludkova A, Stehlik D, Trojanec R, Janostakova A, Korinkova G, Mihal V, Hajduch M. In vitro chemoresistance profile and expression/function of MDR associated proteins in resistant cell lines derived from CCRF-CEM, K562, A549 and MDA MB 231 parental cells. Neoplasma. 2002;49(6):418-25. PMID: 12584592.
Vasan, N., Baselga, J. & Hyman, D.M. A view on drug resistance in cancer. Nature575, 299–309 (2019). https://doi.org/10.1038/s41586-019-1730-1
Galmarini, C., Mackey, J. & Dumontet, C. Nucleoside analogues: mechanisms of drug resistance and reversal strategies. Leukemia15, 875–890 (2001). https://doi.org/10.1038/sj.leu.2402114
Recommended resources
Noskova V, Dzubak P, Kuzmina G, Ludkova A, Stehlik D, Trojanec R, Janostakova A, Korinkova G, Mihal V, Hajduch M. In vitro chemoresistance profile and expression/function of MDR associated proteins in resistant cell lines derived from CCRF-CEM, K562, A549 and MDA MB 231 parental cells. Neoplasma. 2002;49(6):418-25. PMID: 12584592.
Vasan, N., Baselga, J. & Hyman, D.M. A view on drug resistance in cancer. Nature575, 299–309 (2019). https://doi.org/10.1038/s41586-019-1730-1
Galmarini, C., Mackey, J. & Dumontet, C. Nucleoside analogues: mechanisms of drug resistance and reversal strategies. Leukemia15, 875–890 (2001). https://doi.org/10.1038/sj.leu.2402114
Enclosed appendices
-
Appendices bound in thesis
graphs, schemes, tables
Taken from the library
No
Full text of the thesis
Appendices
Reviewer's report
Supervisor's report
Defence procedure record
Obhajobu bakalářské práce studentky Michaely Sapíkové zahájil prof. RNDr. Zdeněk Dvořák, DrSc. et Ph.D. Studentka seznámila komisi s hlavními částmi a výsledky své práce. Následovalo přečtení posudků vedoucí práce RNDr. Lenky Hrubé a oponenta Mgr. Tomáše Oždiana, Ph.D.
Studentka pak reagovala na připomínky a dotazy.
Prezentace studentky obsahovala tato témata:
Rezistence k léčivům, MDR
ABC transportéry
Experimentální provedení práce
V rozpravě byly uvedeny následující připomínky či nastoleny tyto problémy:
Problémy s provedením western blottů
Další experimenty vysvětlující rezistenci léčiv
Způsob psaní bakalářské práce vzhledem ke snadnější orientaci v textu
Doba provádění experimentů vs. žádné zaznamenané zvýšení rezistence vůči testovaným látkám
Různá úroveň cross-rezistence testovaných látek
Rozdíl buněčné linie HCT116 a HCT116 rezistentní vůči 6-thioguaninu
Způsoby normalizace western blottů
Fosforylace proteinů studovaných western blottingem
Studentka na dotazy položené oponentem práce a členy komise reagovala věcně a přiměřeně.