Abstract 3873: Squalene enhanced chemotherapy and radiation therapy sensitivity in mice through the inhibition of DNA damage checkpoint activity

Abstract

Abstract ATM (Ataxia Telangiectasia Mutated) protein kinase plays a crucial role in cellular DNA damage responses. The inhibition of ATM leads to an abolishment of one such signal pathways termed as “checkpoints”. It is expected that the discovery of checkpoint inhibitor will be an effective assistance of anti-cancer therapy. Recent study reported that Wip1, a magnesium dependent protein phosphatase, de-phosphorylates molecules involved DNA damage checkpoint response such ATM (Ser1981). On the other hand, Squalene (SQ) has been thought to assist anti-cancer therapies such as chemo and/or radiation therapy though there is few available data which refer to its detailed mechanisms. We previously showed that SQ enhances the γ-irradiation (IR) sensitivity through the induction of Wip1 in A549 adenocarcinoma cells. It was indicated that IR induced DNA damage checkpoint was abrogated by SQ pre-treatment, and led to the shrinkage of tumor in vivo. Here, we report SQ sensitizes chemotherapy and an advanced radiation therapy, heavy ion (HI) exposure, in mice using allo/xeno-graft model. Mouse Colon-26 and human MKN45 cells (5 × 106 cells/ mouse) were transplanted into Balb/c and SCID mice, respectively. The tumor volumes were estimated by measuring the growing diameters of tumor tissue every day. SQ (200μl, oral administration) was challenged after day-8 and later days. Chemotherapy using DOX (2 mg/kg, i.p.) was conducted in same days with SQ. One-time exposure of HI (Carbon, 290MeV) was performed on the day-8 in NIRS (National Institute of Radiological Sciences, Japan). Marked change of body weight was not observed between the treatments. DOX and HI prevented the tumor growth. SQ itself inhibited tumor growth slightly. The administration of SQ significantly improved the efficacy of DOX and HI on tumor growth. Wip1 protein expression in tumor tissue was detected by western blot analysis, and was elevated by the oral administration of SQ. These results were confirmed in xenograft model using MKN45 cells and SCID mice. Taken together these findings, SQ sensitized chemo/radio therapy through the inhibition of DNA damage checkpoint activity by Wip1. Further in-depth study is required to determine the clinical feasibility of SQ as a sensitizing agent for anti-cancer therapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3873.