Investi̇gati̇on of oncogeni̇c wi̇p1 phosphatase and dna damage response regulati̇on i̇n rhabdomyosarcoma cell li̇ne

Objective: It was aimed to investigate the effect of oncogenic Wip1 phosphatase on genotoxic stress-induced (etoposide) apoptosis activation in RMS A204 cell line. Material and Methods: In the study, using etoposide, GSK2830371, and etoposide + GSK2830371 in A204 cells; Cell viability was tested by MTT analysis after 24-48-72 hours of incubation. Annexin V test was performed at the end of the 24-48-72 hours period to induce apoptosis. The levels of WIP1, GAPDH, phospho p53, and total p53 proteins were measured by Western blot analysis at 48 hours. Results: It has been determined by the MTT test that Etoposide reduces metabolic activity depending on time and dose increase in combination with GSK. It was determined by the Annexin V test that the combination of etoposide and GSK also increased the amount of apoptosis depending on the increase in time. It was determined by Western blot analysis that the combination of etoposide + GSK caused an increase in the level of phospho p53 protein by suppressing Wip1. Conclusion: in this study, the efficacy of etoposide, a chemotherapy agent that creates genotoxic stress by inhibiting the excessively increased expression of Wip1 due to PPM1D amplification in A204 rhabdomyosarcoma cell line, and the response of these cells to chemotherapy were investigated through apoptosis. Key words: DNA damage response (DDR), A204, Rhabdomyosarcoma (RMS), Wip1 phosphatase.