8.–12. 11. 2021
Časové pásmo: Europe/Prague

Proteins of double-strand break and western blot analysis

12. 11. 2021 9:46
Miniprezentace Biologické účinky a zdravotní hlediska Biologické účinky a zdravotní hlediska


Jiří Toufar (Biofyzikální ústav AV ČR, v.v.i.)


Ionising irradiation (IR), such as γ- or X-rays from medical treatments, high energy charged particles from cosmic radiation, is one of most serious factors to endanger human health. IR can produce a variety of DNA damage structures including double-strand breaks (DSB), which are considered to be a most deleterious DNA lesions and could potentially be a major threat to genome stability. Therefore, DSBs need to be addressed and countered by DNA repair pathways and its proteins. This has generated an immense amount of interest in acquiring more knowledge of this topic and created other experimental designs. What we are following is measuring of levels of various proteins in response to γ- irradiation of different cell lines – U87 (human primary glioblastoma cell line), SkBr3 (human breast cancer cell line) and NHDF (Normal Human Dermal Fibroblasts). Using this, we try to find out the selected pathway for DNA repair and monitor this at different times after IR. Western blotting analysis is there used for determining levels of γH2AX, MRE11, RAD51, PARP1, 53BP1 and other proteins of interest occurring in separate DNA damage response mechanisms. First results show promise and indicate us some variances in protein levels in different times after IR and more importantly some similar patterns. In future plans, we will try to use different types of nanoparticles and observe how protein levels are affected after γ or possibly X-ray irradiation and moreover, if the chosen pathway of DNA repair mechanism is modified or changed.

Supported by the project: DFG 20-04109 J

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Hlavní autoři

Jiří Toufar (Biofyzikální ústav AV ČR, v.v.i.) Lucie Dobešová (Biofyzikální ústav AV ČR, v.v.i.) Eva Pagáčová (Biofyzikální ústav AV ČR, v.v.i.) Olga Kopečná (Biofyzikální ústav AV ČR, v.v.i.) Iva Falková (Biofyzikální ústav AV ČR, v.v.i.) Alena Bačíková (Biofyzikální ústav AV ČR, v.v.i.) Michael Hausmann (Kirchhoff-Institute for Physics, University of Heidelberg) Martin Falk (Biofyzikální ústav AV ČR, v.v.i.)

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