Abstract 885: Induction of PRDX2 by H. pylori reduces ROS and promotes cancer cell survival and resistance to cisplatin

Abstract

Abstract Background & Aims: The antioxidant enzyme peroxiredoxin 2 (PRDX2) plays a critical role in regulating reactive oxygen species (ROS) levels in several diseases. Helicobacter pylori (H. pylori) infection is a well-known risk factor of gastric cancer. The role of PRDX2 in gastric tumorigenesis remains largely unknown. We investigated the molecular function and regulation of PRDX2 in response to infection with H. pylori and cisplatin treatment in gastric cancer cells. Methods:Western blots (WB) and quantitative real-time PCR (qPCR) analysis were performed on AGS, SNU-1 and MKN28 gastric cancer cell lines to detect PRDX2 expression levels with or without H. pyloriinfection (7.13 or J166 strain). We evaluated the levels of ROS by H2DCFDA staining. WB analysis was used to determine oxidative DNA damage and double stranded DNA breaks by using antibodies against 8-Oxo-guanine and p-H2AX. WB and NF-κb luciferase reporter assay were performed to investigate the link between PRDX2 and NF-κb signaling. ATP-GLO cell titer analysis was utilized to determine cell viability. Results:Our data indicated that PRDX2 mRNA and protein expression levels were induced byH. pyloriin AGS and SNU-1 cells. PRDX2 knockdown significantly increased ROS levels and 8-Oxoguanine staining followingH. pyloriinfection in both cells. These results were further confirmed by Western blot data showing that p-H2AX protein level was strongly induced in PRDX2 knockdown and H. pyloriinfected cells. Interestingly, Western blot data showed that TNF-αtreatment induced PRDX2 protein levels while Bay 11-7082 treatment decreased PRDX2 protein level in AGS and SNU-1 cells. At the same time, PRDX2 transient knockdown in both AGS and SNU-1 cells decreased p-P65 (S536) protein expression levels, nuclear localization of NF-κB-p65, and luciferase reporter activity (P<0.05). We also found that inhibition of PRDX2 significantly sensitized AGS and SNU-1 cells to cisplatin treatment. Conclusion:Our data indicates that knockdown of PRDX2 enhances ROS and DNA damage with H. pylori infection on gastric cancer cells after H. pylori infection and sensitizes gastric cancer cells to cisplatin treatment. Our results suggest a positive feedforward loop between PRDX2 and NF-kB to reduce ROS levels. KEY WORDS: PRDX2,H. pylori, gastric cancer, reactive oxygen species, DNA damage, NF-κb Citation Format: Sen Wang, Zheng Chen, Heng Lu, Shoumin Zhu, Dunfa Peng, Mohammed Soutto, Ahmed Gomma, Nadeem Bhat, Huma Naz, Zekuan Xu, Wael El-Rifai. Induction of PRDX2 by H. pylori reduces ROS and promotes cancer cell survival and resistance to cisplatin [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 885.