Andersson Maria A, Hellman Björn E
Department of Pharmaceutical Biosciences, Division of Toxicology, BMC, SE-751 24 Uppsala, Sweden.
Toxicol In Vitro. 2007 Jun;21(4):716-22. doi: 10.1016/j.tiv.2006.12.012. Epub 2007 Jan 10.
Extended-term cultures of proliferating human T-lymphocytes (ETC) may be a practical alternative to freshly isolated non-proliferating peripheral blood lymphocytes (PBL) when studying genotoxicity in vitro. To investigate if the pattern of DNA damage differs between the two in vitro systems, catechol-induced DNA damage was evaluated in PBL and ETC derived from the same blood sample, using three different donors. DNA damage was monitored using the comet assay. Whereas 3 h of exposure to 0.5 mM catechol was found to be without DNA damaging effects, 3 mM was found to induce significant damage both in the PBL and the ETC (the latter being clearly less sensitive). The level of reactive oxygen species (ROS) was also measured in the ETC using the fluorescent probe carboxy-H2DCFA. ROS was found to be considerably increased both at 0.5 and 3 mM catechol. The demonstrated difference in sensitivity towards catechol-induced DNA damage between PBL and ETC may be due to their different proliferative status, but despite this difference both in vitro systems were able to identify catechol as a DNA damaging agent at the same concentration.
在体外研究遗传毒性时，增殖性人类T淋巴细胞的长期培养物（ETC）可能是新鲜分离的非增殖外周血淋巴细胞（PBL）的一种实用替代物。为了研究这两种体外系统中DNA损伤模式是否不同，使用来自三个不同供体的同一血样，对PBL和ETC中儿茶酚诱导的DNA损伤进行了评估。使用彗星试验监测DNA损伤。虽然发现暴露于0.5 mM儿茶酚3小时没有DNA损伤作用，但发现3 mM儿茶酚在PBL和ETC中均诱导了显著损伤（后者明显不那么敏感）。还使用荧光探针羧基-H2DCFA在ETC中测量了活性氧（ROS）水平。发现在0.5 mM和3 mM儿茶酚时ROS均显著增加。PBL和ETC对儿茶酚诱导的DNA损伤的敏感性差异可能是由于它们不同的增殖状态，但尽管存在这种差异，两种体外系统都能够在相同浓度下将儿茶酚鉴定为DNA损伤剂。
