Methoxyamine potentiates iododeoxyuridine-induced radiosensitization by altering cell cycle kinetics and enhancing senescence

• Published in: Molecular cancer therapeutics 2006-04, Vol.5 (4), p.893-902
• Main Authors: Yan, Tao, Seo, Yuji, Schupp, Jane E, Zeng, Xuehuo, Desai, Anand B, Kinsella, Timothy J
• Format: Article
• Language: English
• Subjects: Bromodeoxyuridine - pharmacology; cell cycle; Cell Cycle - drug effects; Cell Cycle - radiation effects; Cell Line, Tumor; Cell Survival - drug effects; Cell Survival - radiation effects; Cellular Senescence - drug effects; Colorectal Neoplasms; G1 Phase - drug effects; G1 Phase - radiation effects; Humans; Hydroxylamines - pharmacology; Idoxuridine - pharmacology; ionizing radiation; IUdR; Kinetics; methoxyamine; Radiation-Sensitizing Agents - pharmacology; senescence
• ISSN: 1535-7163; 1538-8514
• DOI: 10.1158/1535-7163.MCT-05-0364

We previously reported that methoxyamine (an inhibitor of base excision repair) potentiates iododeoxyuridine (IUdR)–induced radiosensitization in human tumor cells. In this study, we investigated the potential mechanisms of this enhanced cell death. Human colorectal carcinoma RKO cells were exposed to IUdR (3 μmol/L) and/or methoxyamine (3 mmol/L) for 48 hours before ionizing radiation (5 Gy). We found that IUdR/methoxyamine altered cell cycle kinetics and led to an increased G 1 population but a decreased S population before ionizing radiation. Immediately following ionizing radiation (up to 6 hours), IUdR/methoxyamine–pretreated cells showed a stringent G 1 -S checkpoint but an insufficient G 2 -M checkpoint, whereas a prolonged G 1 arrest, containing 2CG1 and 4CG1 cells, was found at later times up to 72 hours. Levels of cell cycle–specific markers [p21, p27, cyclin A, cyclin B1, and pcdc2(Y15)] and DNA damage signaling proteins [γH2AX, pChk1(S317), and pChk2(T68)] supported these altered cell cycle kinetics. Interestingly, we found that IUdR/methoxyamine pretreatment reduced ionizing radiation–induced apoptosis. Additionally, the extent of cell death through necrosis or autophagy seemed similar in all (IUdR ± methoxyamine + ionizing radiation) treatment groups. However, a larger population of senescence-activated β-galactosidase-positive cells was seen in IUdR/methoxyamine/ionizing radiation–treated cells, which was correlated with the increased activation of the senescence factors p53 and pRb. These data indicate that IUdR/methoxyamine pretreatment enhanced the effects of ionizing radiation by causing a prolonged G 1 cell cycle arrest and by promoting stress-induced premature senescence. Thus, senescence, a novel ionizing radiation–induced tumor suppression pathway, may be effectively targeted by IUdR/methoxyamine pretreatment, resulting in an improved therapeutic gain for ionizing radiation. [Mol Cancer Ther 2006;5(4):893–902]