Genotoxicity of aristolochic acid in human cell lines

• Main Authors: Yu-Yun Wei, 魏玉雲
• Other Authors: Wushou P. Chang
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/66617783032372637791

碩士 === 國立陽明大學 === 環境衛生研究所 === 91 === 　Aristolochic acid (AA), derived from Aristolochia spp., was related to the development of a novel nephropathy and urothelial carcinomas in patients with aristolochic acid nephropathy (AAN). Since treatment of mammalian cells with AA—a mixture of AAI and AAII─has been shown to result in induction of DNA mutagenesis and carcinoma in animals, this study is designed to investigate whether genotoxicity can be induced in AA-treated human skin fibroblast (HSF) and 1BR3 cells. After various AA exposure, HSF and 1BR3 cells were evaluated for the frequencies of nuclear foci by labeling with anti g-H2AX monoclonal antibody, which can detect repair proteins for DNA double-strand breaks, as well as micronuclei and p53 protein. 　Exposure of AA led to significant decrease in viabilities of both HSF and 1BR3 cells in dose- and time-dependent manners. Slight time-dependent, but not dose-dependent, increases in proportions of g-H2AX nuclear foci in both HSF and 1BR3 cells were shown after AA exposure. With the frequencies of g-H2AX nuclear foci are 0.77-1.77% in HSF cells and 0.58-2.31% in 1BR3 cells. Dose-dependent increase in the micronuclei frequencies (MF) was shown in HSF cells. MF in the binucleated HSF cells induced by cytochalasin-B was higher than those observed in mononucleated ones. HSF cells post AA was shown with increase in total p53 proteins. Furthermore, the levels of p53 were found to be dose- and time-dependent in HSF cells after AA exposure. There was a high association between MF and p53 proteins (R>0.84) in the dose-dependent manner. These indicated that AA is with genotoxic and/or mutagenic effects in human cells, and both MF and p53 protein were more sensitive for the evaluation. The association between DNA adducts and other DNA damages induced by AA in human cells remains to be further established.