Błasiak J, Sikora A, Wozniak K, Drzewoski J
Department of Molecular Genetics, University of Lodz, Lodz, Poland.
Cell Biol Toxicol. 2004 Mar;20(2):83-96. doi: 10.1023/b:cbto.0000027919.38379.28.
Streptozotocin (STZ) is an antibiotic which can be used to induce diabetes in experimental animals in order to have an insight into pathogenesis of this disease. To use STZ as a diabetogenic substance, its molecular mode of action should be elucidated. Using the alkaline comet assay, we showed that STZ at concentrations in the range 0.01-100 micromol/L induced DNA damage in normal human lymphocytes and HeLa cancer cells in a dose-dependent manner. Lymphocytes were able to remove damage to their DNA within a 30-min repair incubation, whereas HeLa cells completed the repair in 60 min. Vitamins C and E at 10 and 50 micromol/L diminished the extent of DNA damage induced by 50 micromol/L STZ. Pretreatment of the lymphocytes with the nitrone spin trap, alpha-(4-pyridil-1-oxide)-N-tert-butylnitrone (POBN) or ebselen, which mimics glutathione peroxidase, or pyrrolidine dithiocarbamate (PDTC) reduced the extent of DNA damage evoked by STZ. The cells exposed to STZ and treated with endonuclease III (Endo III), formamidopyrimidine-DNA glycosylase (Fpg) and 3-methyladenine-DNA glycosylase II (AlkA), the enzymes recognizing oxidized and alkylated bases, displayed greater extent of DNA damage than those not treated with these enzymes. These results suggest that free radicals may be involved in the formation of DNA lesions induced by streptozotocin. The drug can also alkylate DNA bases. This broad range of DNA damage induced by STZ indicates that the drug may seriously affect genomic stability in normal and pathological cells.
链脲佐菌素(STZ)是一种抗生素,可用于在实验动物中诱导糖尿病,以便深入了解该疾病的发病机制。为了将STZ用作致糖尿病物质,应阐明其分子作用模式。使用碱性彗星试验,我们发现浓度在0.01 - 100微摩尔/升范围内的STZ以剂量依赖方式诱导正常人淋巴细胞和HeLa癌细胞的DNA损伤。淋巴细胞能够在30分钟的修复孵育内修复其DNA损伤,而HeLa细胞在60分钟内完成修复。10和50微摩尔/升的维生素C和E可减少50微摩尔/升STZ诱导的DNA损伤程度。用硝酮自旋捕获剂α -(4 - 吡啶基 - 1 - 氧化物)- N - 叔丁基硝酮(POBN)或模拟谷胱甘肽过氧化物酶的依布硒仑或吡咯烷二硫代氨基甲酸盐(PDTC)预处理淋巴细胞可降低STZ引起的DNA损伤程度。暴露于STZ并用识别氧化和烷基化碱基的酶——内切酶III(Endo III)、甲酰胺嘧啶 - DNA糖基化酶(Fpg)和3 - 甲基腺嘌呤 - DNA糖基化酶II(AlkA)处理的细胞,比未用这些酶处理的细胞显示出更大程度的DNA损伤。这些结果表明自由基可能参与链脲佐菌素诱导的DNA损伤形成。该药物还可使DNA碱基烷基化。STZ诱导的这种广泛的DNA损伤表明该药物可能严重影响正常和病理细胞中的基因组稳定性。