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基于质谱的方法来测量甲毒素B1 在甲素固定的胺嵌入组织中的DNA附加物.

Medjda Bellamri, Lihua Yao, Rachana Tomar1

  • 1Department of Chemistry, Vanderbilt Ingram Cancer Center, Vanderbilt University, Nashville, Tennessee 37235, United States.

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甲素B1 (AFB1) DNA添加物,AFB1-FapyGua,在甲固定组织中是稳定的. 这一突破允许在人类肝脏样本中对AFB1暴露的生物监测,有助于对这种强有力的致癌物进行研究.

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科学领域:

  • 毒理学 毒理学 毒理学
  • 分子生物学分子生物学
  • 癌症研究 癌症研究

背景情况:

  • 非洲毒素B1 (AFB1) 是一种由菌产生的强有力的人类肝脏致癌物,污染了主食.
  • AFB1生物激活形成DNA添加物,包括AFB1-N7-瓜 (AFB1-N7-Gua) 和环开放的AFB1-FapyGua添加物.
  • AFB1-FapyGua添加物与AFB1-诱导的G → T突变和肝癌有关,但它们在人类中的检测受到组织可用性的限制.

研究的目的:

  • 为了评估AFB1-FapyGua添加物的稳定性,在甲固定式嵌 (FFPE) 组织中.
  • 建立一种检测FFPE人类肝脏样本中的AFB1-FapyGua添加物的方法,用于生物监测.
  • 克服与甲固定相关的核酸质量挑战,以准确量化添加物.

主要方法:

  • 新生小鼠接触AFB的情况1.
  • 使用离子陷和轨道陷质谱仪在新鲜冷和FFPE肝脏组织中分析AFB1-FapyGua附加物.
  • 开发FFPE样本的DNA去交叉链接检索流程.

主要成果:

  • 环开放的AFB1-FapyGua添加物在甲固定和DNA脱交联过程中是稳定的.
  • 在FFPE肝脏中检测到AFB1-FapyGua附加物,其水平与新鲜冷肝脏相当.
  • 轨道飞行器MS2的量化极限为每10^8个基中有4.0个基,含有0.8μg的DNA.

结论:

  • 在FFPE肝组织中,AFB1-FapyGua附加物可靠地检测到.
  • 这种DNA检索技术可以在有风险的人群中对AFB1暴露进行生物监测.
  • 这些发现促进了使用可访问的FFPE样本研究AFB1相关的肝癌发生.