Cookies on this website
We use cookies to ensure that we give you the best experience on our website. If you click 'Continue' we'll assume that you are happy to receive all cookies and you won't see this message again. Click 'Find out more' for information on how to change your cookie settings.
Skip to main content

Colony formation is the gold standard assay for determining reproductive cell death after radiation treatment, since effects on proliferation often do not reflect survival. We have developed a high-throughput radiosensitivity screening method based on clonogenicity and screened a siRNA library against kinases. Thiamine pyrophosphokinase-1 (TPK1), a key component of Vitamin B1/thiamine metabolism, was identified as a target for radiosensitization. TPK1 knockdown caused significant radiosensitization in cancer but not normal tissue cell lines. Other means of blocking this pathway, knockdown of thiamine transporter-1 (THTR1) or treatment with the thiamine analogue pyrithiamine hydrobromide (PyrH) caused significant tumor specific radiosensitization. There was persistent DNA damage in cells irradiated after TPK1 and THTR1 knockdown or PyrH treatment. Thus this screen allowed the identification of thiamine metabolism as a novel radiosensitization target that affects DNA repair. Short-term modulation of thiamine metabolism could be a clinically exploitable strategy to achieve tumor specific radiosensitization.

Original publication

DOI

10.18632/oncotarget.3468

Type

Journal article

Journal

Oncotarget

Publication Date

20/03/2015

Volume

6

Pages

5978 - 5989

Keywords

TPK1, high-throughput screening, thiamine, tumor radiosensitivity, Cell Line, Tumor, Colony-Forming Units Assay, DNA Damage, HCT116 Cells, HeLa Cells, High-Throughput Screening Assays, Humans, Neoplasms, Pyrithiamine, Radiation Tolerance, Radiation-Sensitizing Agents, Thiamin Pyrophosphokinase, Thiamine, Transfection