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首页> 外文期刊>Mutation Research, C. Mutation Research Letters >PREFERENTIAL BINDING OF CISPLATIN TO MITOCHONDRIAL DNA OF CHINESE HAMSTER OVARY
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PREFERENTIAL BINDING OF CISPLATIN TO MITOCHONDRIAL DNA OF CHINESE HAMSTER OVARY

机译：喜树碱与中国仓鼠卵巢线粒体DNA的优先结合

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摘要

Some chemical carcinogens localize preferentially in mitochondrial DNA (mtDNA) when compared with genomic DNA (gDNA). Here we compare the ability of cisplatin (cis-diamminedichloroplatinum[II]) to induce DNA adducts in both genomic and mtDNA of Chinese hamster ovary (CHO) cells in culture. Cytotoxicity was examined by cell survival 4, 8 and 24 h after exposure to 50 mu M cisplatin. Cisplatin-DNA adducts were measured in DNA from nuclear and mitochondrial fractions by dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA), a sensitive competitive microtiter-based immunoassay utilizing antiserum elicited against cisplatin-modified DNA. An additional comparison of cisplatin-DNA binding in both compartments was performed by immunoelectron microscopy using the cisplatin-DNA antiserum and colloidal gold. DELFIA analysis of cisplatin-DNA adducts in gDNA and mtDNA showed a six-fold higher incorporation of drug into mtDNA as compared to gDNA. Morphometric studies of colloidal gold distribution in photomicrographs of CHO cells showed mtDNA to contain a four-fold higher concentration of cisplatin as compared to nuclear DNA. Therefore, both methods demonstrated a preferential binding of cisplatin to mtDNA versus gDNA.
机译：与基因组DNA（gDNA）相比，某些化学致癌物优先位于线粒体DNA（mtDNA）中。在这里，我们比较了顺铂（cis-diamminedichloroplatinum [II]）诱导培养的中国仓鼠卵巢（CHO）细胞基因组和mtDNA中的DNA加合物的能力。通过暴露于50μM顺铂后4、8和24小时的细胞存活来检查细胞毒性。通过离解增强型镧系元素荧光免疫测定法（DELFIA）测定了核和线粒体级分的DNA中的顺铂DNA加合物，这是一种敏感的基于竞争性微量滴定的免疫测定法，利用抗血清对顺铂修饰的DNA进行诱导。通过使用顺铂-DNA抗血清和胶体金的免疫电子显微镜对两个隔室中的顺铂-DNA结合进行了另外的比较。 DELFIA分析gDNA和mtDNA中的顺铂-DNA加合物显示，与gDNA相比，将药物掺入mtDNA的效率高六倍。 CHO细胞的显微照片中胶体金分布的形态计量学研究表明，与核DNA相比，mtDNA包含的顺铂浓度高四倍。因此，两种方法都证明了顺铂相对于mtDNA相对于gDNA的优先结合。

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