DNA damage sensitization of breast cancer cells with PARP10/ARTD10 inhibitor

• Main Author: Hukkanen, M. (Mikko)
• Format: Dissertation
• Language: English
• Published: University of Oulu 2019
• Online Access: http://jultika.oulu.fi/Record/nbnfioulu-201909062843

Abstract. In this thesis, I studied the DNA damage sensitization of breast cancer cells with PARP10/ARTD10 inhibitor. As the ARTD10 inhibition field is relatively fresh, tests with breast cancer cell lines combined with clinically used chemotherapeutics will elucidate the potential future uses of the inhibitors in a clinically relevant context, directing the future research efforts in the field. To study the link between OUL35 and DNA damage sensitization, cell proliferation experiments were conducted, and to determine whether ARTD10 translocation from cytoplasm into nucleus is enhanced under DNA damaging conditions, western blot assay was performed. Also, the OUL35 potency against full-length ARTD10 was verified to compare it with the catalytic ARTD10 fragment. The determined IC50 of 510 nM suggests that the potency does not differ from studies with catalytic domain. According to the results from ARTD10 translocation studies, it can only be said that DNA damaging agent in general reduces the amount of cytoplasmic ARTD10, whereas I could not confirm the potential translocation to the nucleus. The results of OUL35-mediated sensitization to DNA damaging agents indicates that OUL35 might sensitize breast cancer cells to DNA damage. As a summary, full-length ARTD10 inhibition does not vary from catalytic domain, suggesting that either construct can be used for testing inhibitors, OUL35 may enhance the effect of other DNA damaging agents, and there is a possibility that HU might have an influence on the nuclear translocation of ARTD10. This provides the basis for future evaluation of larger cancer cell panels for sensitization for chemotherapeutics by ARTD10 inhibition.