Characterization of oncogenic role of ROR1 and PTK7 in B-ALL
Hautanen, Veera (2018)
2018
Bioteknologian tutkinto-ohjelma - Degree Programme in Biotechnology
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Hyväksymispäivämäärä
2018-10-12
Julkaisun pysyvä osoite on
http://urn.fi/URN:NBN:fi:uta-201810192725
http://urn.fi/URN:NBN:fi:uta-201810192725
Tiivistelmä
Background and aims: Pseudokinases are members of the protein kinase superfamily that have mutations in their canonical motifs required for proper catalytic activity. Many pseudokinases are involved in human diseases, therefore it is important to understand their signaling mechanisms and regulation. Among pseudokinases family, receptor tyrosine kinase-like orphan receptors (RORs) and protein tyrosine kinase 7 (PTK7) are Wnt-binding receptors from non-canonical Wnt signaling pathway. These proteins are expressed in several cancers, but their functional role needs more investigation. In this thesis, we investigated the oncogenic role of ROR1 and PTK7 in B-cell acute lymphoblastic leukemia (B-ALL).
Methods: In this study, we used B-ALL cells with t(1;19) translocation. We investigated the expression of ROR1 and PTK7 as well as intracellular Wnt signaling pathway using Western Blotting and FACS analysis. We also examined the interaction of Wnt5a ligand with ROR1 and PTK7 using immunoprecipitations. To examine molecular pathway downstream from ROR1 and the relevance of ROR1 and PTK7 in cell survival, we targeted ROR1 with monoclonal antibody and both ROR1 and PTK7 with RNA interference system.
Results: Our results demonstrate that both ROR1 an PTK7 are expressed on B-ALL cells with t(1;19) translocation but only ROR1 is important for B-ALL cell survival. We showed that ROR1 interacts with Wnt5a indicating that ROR1 is a mediator of non-canonical Wnt signaling. Also, we demonstrated downregulation of RhoA, STAT3 and NF-?B after ROR1 targeting.
Conclusion: ROR1 has an important role maintaining B-ALL t(1;19) cell viability in non-canonical Wnt signaling pathway. ROR1 targeting reduces cell survival indicating it as a possible candidate for targeted therapy.
Methods: In this study, we used B-ALL cells with t(1;19) translocation. We investigated the expression of ROR1 and PTK7 as well as intracellular Wnt signaling pathway using Western Blotting and FACS analysis. We also examined the interaction of Wnt5a ligand with ROR1 and PTK7 using immunoprecipitations. To examine molecular pathway downstream from ROR1 and the relevance of ROR1 and PTK7 in cell survival, we targeted ROR1 with monoclonal antibody and both ROR1 and PTK7 with RNA interference system.
Results: Our results demonstrate that both ROR1 an PTK7 are expressed on B-ALL cells with t(1;19) translocation but only ROR1 is important for B-ALL cell survival. We showed that ROR1 interacts with Wnt5a indicating that ROR1 is a mediator of non-canonical Wnt signaling. Also, we demonstrated downregulation of RhoA, STAT3 and NF-?B after ROR1 targeting.
Conclusion: ROR1 has an important role maintaining B-ALL t(1;19) cell viability in non-canonical Wnt signaling pathway. ROR1 targeting reduces cell survival indicating it as a possible candidate for targeted therapy.