From the Department of Nutrition and Food Science, University of Maryland, College Park, Maryland 20742; School of Life Science, Lanzhou University, Lanzhou, Gansu 730000, China.
Diet, Genomics, and Immunology Laboratory, Beltsville Human Nutrition Research Center, United States Department of Agriculture, Beltsville, Maryland 20705.
J Biol Chem. 2010 Oct 22;285(43):33010-33017. doi: 10.1074/jbc.M110.137406. Epub 2010 Aug 6.
Epidemiological and animal studies indicate that selenium supplementation suppresses risk of colorectal and other cancers. The majority of colorectal cancers are characterized by a defective DNA mismatch repair (MMR). Here, we have employed the MMR-deficient HCT 116 colorectal cancer cells and the MMR-proficient HCT 116 cells with hMLH1 complementation to investigate the role of hMLH1 in selenium-induced DNA damage response, a tumorigenesis barrier. The ATM (ataxia telangiectasia mutated) protein responds to clastogens and initiates DNA damage response. We show that hMLH1 complementation sensitizes HCT 116 cells to methylseleninic acid, methylselenocysteine, and sodium selenite via reactive oxygen species and facilitates the selenium-induced oxidative 8-oxoguanine damage, DNA breaks, G(2)/M checkpoint response, and ATM pathway activation. Pretreatment of the hMLH1-complemented HCT 116 cells with the antioxidant N-acetylcysteine or 2,2,6,6-tetramethylpiperidine-1-oxyl or the ATM kinase inhibitor KU55933 suppresses hMLH1-dependent DNA damage response to selenium exposure. Selenium treatment stimulates the association between hMLH1 and hPMS2 proteins, a heterodimer critical for functional MMR, in a manner dependent on ATM and reactive oxygen species. Taken together, the results suggest a new role of selenium in mitigating tumorigenesis by targeting the MMR pathway, whereby the lack of hMLH1 renders the HCT 116 colorectal cancer cells resistant to selenium-induced DNA damage response.
流行病学和动物研究表明,硒补充剂可降低结直肠癌和其他癌症的风险。大多数结直肠癌的特征是 DNA 错配修复 (MMR) 缺陷。在这里,我们使用 MMR 缺陷的 HCT 116 结直肠癌细胞和 MMR 功能正常的 HCT 116 细胞与 hMLH1 互补来研究 hMLH1 在硒诱导的 DNA 损伤反应中的作用,这是一种肿瘤发生障碍。ATM(共济失调毛细血管扩张突变)蛋白对断裂剂作出反应并启动 DNA 损伤反应。我们表明,hMLH1 互补使 HCT 116 细胞对甲基硒酸、甲基硒代半胱氨酸和亚硒酸钠敏感,通过活性氧和促进硒诱导的氧化 8-氧鸟嘌呤损伤、DNA 断裂、G2/M 检查点反应和 ATM 途径激活。用抗氧化剂 N-乙酰半胱氨酸或 2,2,6,6-四甲基哌啶-1-氧自由基或 ATM 激酶抑制剂 KU55933 预处理 hMLH1 互补的 HCT 116 细胞会抑制硒暴露后 hMLH1 依赖性 DNA 损伤反应。硒处理以依赖于 ATM 和活性氧的方式刺激 hMLH1 与 hPMS2 蛋白之间的关联,hPMS2 蛋白是功能性 MMR 的关键异二聚体。总之,这些结果表明,硒通过靶向 MMR 途径在减轻肿瘤发生方面发挥了新的作用,其中 hMLH1 的缺失使 HCT 116 结直肠癌细胞对硒诱导的 DNA 损伤反应产生抗性。