Within this PhD proposal, we will investigate the impact of different radiation qualities on Hedgehog signaling in cancer cells.
More specific, we will:
(1) further investigate the cellular and molecular response to low and high doses of different radiation qualities
(2) study how Hh signaling inhibitors can modulate the sensitivity of cancer cells to low and high doses of ionizing radiation of different qualities.
All experiments will be performed using in vitro human cancer cell lines, more specific a prostate cancer cell line (PC3) and a medulloblastoma cell line (Daoy- will be used. Not only are both cancer type indications for the use of particle therapy, but the Hh signaling pathway seems to play an important role both in the initiation as well as the progression and metastasis of these cancers.
Low-LET X-ray irradiation will be performed at the irradiation facility of SCK•CEN. For irradiation experiments with high-LET beams, we will continue to apply for beam time to different European accelerator facilities including GSI (Darmstadt, Germany), GANIL (Caen, France), and LNS-INFN (Catania, Italy).
To investigate the cellular and molecular response of these cancer cells to low and high doses of different radiation qualities, several experiments will be done: a) analysis of changes in gene expression involved in the Hh pathway and migration pathways, b) cell migration and motility assays to examine the potential changes in cell behavior, and c) colony formation assays to determine clonogenic cell survival.
In the second part of this PhD project, the cancer cell lines will be treated with different Hh signaling inhibitors and their effect on cell survival and cell behavior after irradiation with different radiation qualities will be investigated by means of clonogenic assays and cell migration/motility assays respectively.