Establishment of a Transformation Coupled in vitro End Joining Assay to Estimate Radiosensitivity in Tumor Cells

Establishment of a Transformation Coupled in vitro End Joining Assay to Estimate Radiosensitivity in Tumor Cells

Blog Article

Here, we present a modified in vitro end-joining (EJ) assay to quantify EJ capacity, accuracy as well as pathway switch to alternative end-joining (Alt-EJ) or single strand annealing (SSA).A novel transformation assay was established to specifically measure circular repair products, which correlate with classical EJ efficiency.The EJ assay was validated using EJ-deficient mammalian cell lines (Ku80, DNA-PKcs, LigIV, or XRCC4 mutants).

A pathway switch to Alt-EJ and SSA was seen Car Model Kit exclusively in Ku-deficient cells.Circular EJ product formation correlated with cell survival and DSB repair capacity after X-irradiation.Investigation of 14 HNSCC cell lines revealed differences in the total EJ capacity but a broader variation in the amount of circular repair products.

Sequencing of repair junctions in HNSCC cells demonstrated a predominance of high-fidelity EJ and an avoidance of both Alt-EJ and SSA.A significant correlation was observed between the amount of circular repair products, repair of IR-induced DSB and radiosensitivity.Collectively, these data indicate that the presented in vitro-EJ-assay can not only estimate the repair capacity Airliner Model Kits of cancer cells to enable the stratification into radiosensitive or radioresistant, but can also identify repair pathway deregulation such as a switch to Alt-EJ or SSA, which enables tumor targeting.