Division of Chest Medicine, Department of Internal Medicine, Changhua Christian Hospital, Changhua 500, Taiwan, ROC.
Oncol Rep. 2012 Apr;27(4):959-64. doi: 10.3892/or.2012.1622. Epub 2012 Jan 4.
It has been shown that deguelin, one of the compounds of rotenoids from flavonoid family, induced cytotoxic effects through induction of cell cycle arrest and apoptosis in many types of human cancer cell lines, but deguelin-affected DNA damage and repair gene expression (mRNA) are not clarified yet. We investigated the effects of deguelin on DNA damage and associated gene expression in human lung cancer NCI-H460 cells in vitro. DNA damage was assayed by using the comet assay and DNA gel electrophoresis and the results indicated that NCI-H460 cells treated with 0, 50, 250 and 500 nM deguelin led to a longer DNA migration smear based on the single cell electrophoresis and DNA fragmentation occurred based on the examination of DNA gel electrophoresis. DNA damage and repair gene expression (mRNA) were evaluated by using real-time PCR assay and the results indicated that 50 and 250 nM deguelin for a 24-h exposure in NCI-H460 cells, decreased the gene levels of breast cancer 1, early onset (BRCA1), DNA-dependent serine/threonine protein kinase (DNA-PK), O6-methylguanine-DNA methyltransferase (MGMT), p53, ataxia telangiectasia mutated (ATM) and ataxia-telangiectasia and Rad3-related (ATR) mRNA expressions. Collectively, the present study showed that deguelin caused DNA damage and inhibited DNA damage and repair gene expressions, which might be due to deguelin-inhibited cell growth in vitro.
已证明，脱乙酰基醉椒素（deguelin）是类黄酮家族中的植物固醇化合物之一，它可通过诱导细胞周期停滞和细胞凋亡，对多种人类癌细胞系产生细胞毒性作用，但脱乙酰基醉椒素影响 DNA 损伤和修复基因表达（mRNA）的机制尚不清楚。我们研究了脱乙酰基醉椒素对体外人肺癌 NCI-H460 细胞的 DNA 损伤和相关基因表达的影响。通过彗星试验和 DNA 凝胶电泳检测 DNA 损伤，结果表明，NCI-H460 细胞用 0、50、250 和 500 nM 脱乙酰基醉椒素处理后，基于单细胞电泳的 DNA 迁移出现更长的电泳带，基于 DNA 凝胶电泳的 DNA 片段化发生。通过实时 PCR 检测 DNA 损伤和修复基因表达（mRNA），结果表明，NCI-H460 细胞暴露于 50 和 250 nM 脱乙酰基醉椒素 24 小时后，乳腺癌 1 基因（BRCA1）、DNA 依赖性丝氨酸/苏氨酸蛋白激酶（DNA-PK）、O6-甲基鸟嘌呤-DNA 甲基转移酶（MGMT）、p53、共济失调毛细血管扩张突变（ATM）和共济失调毛细血管扩张和 Rad3 相关（ATR）mRNA 的基因水平降低。综上所述，本研究表明脱乙酰基醉椒素可引起 DNA 损伤，并抑制 DNA 损伤和修复基因表达，这可能是由于脱乙酰基醉椒素抑制了细胞在体外的生长。
