Departments of Health and Nutrition Biotechnology, Asia University, Taichung 413, Taiwan, ROC.
Oncol Rep. 2013 Feb;29(2):613-8. doi: 10.3892/or.2012.2170. Epub 2012 Dec 7.
Numerous studies have demonstrated that triptolide induces cell cycle arrest and apoptosis in human cancer cell lines. However, triptolide-induced DNA damage and inhibition of DNA repair gene expression in human skin cancer cells has not previously been reported. We sought the effects of triptolide on DNA damage and associated gene expression in A375.S2 human malignant melanoma cells in vitro. Comet assay, DAPI staining and DNA gel electrophoresis were used for examining DNA damage and results indicated that triptolide induced a longer DNA migration smear based on single cell electrophoresis and DNA condensation and damage occurred based on the examination of DAPI straining and DNA gel electrophoresis. The real-time PCR technique was used to examine DNA damage and repair gene expression (mRNA) and results indicated that triptolide led to a decrease in the ataxia telangiectasia mutated (ATM), ataxia-telangiectasia and Rad3-related (ATR), breast cancer 1, early onset (BRCA-1), p53, DNA-dependent serine/threonine protein kinase (DNA-PK) and O6-methylguanine-DNA methyltransferase (MGMT) mRNA expression. Thus, these observations indicated that triptolide induced DNA damage and inhibited DNA damage and repair-associated gene expression (mRNA) that may be factors for triptolide-mediated inhibition of cell growth in vitro in A375.S2 cells.
大量研究表明，雷公藤红素可诱导人癌细胞系的细胞周期停滞和凋亡。然而，雷公藤红素诱导人皮肤癌细胞中的 DNA 损伤和抑制 DNA 修复基因表达尚未见报道。我们研究了雷公藤红素对体外 A375.S2 人恶性黑素瘤细胞中 DNA 损伤及相关基因表达的影响。彗星试验、DAPI 染色和 DNA 凝胶电泳用于检测 DNA 损伤，结果表明雷公藤红素诱导基于单细胞电泳的更长的 DNA 迁移拖尾，并且基于 DAPI 染色和 DNA 凝胶电泳的检查，发生了 DNA 浓缩和损伤。实时 PCR 技术用于检测 DNA 损伤和修复基因表达（mRNA），结果表明雷公藤红素导致共济失调毛细血管扩张突变（ATM）、共济失调毛细血管扩张和 Rad3 相关（ATR）、乳腺癌 1，早发（BRCA-1）、p53、DNA 依赖性丝氨酸/苏氨酸蛋白激酶（DNA-PK）和 O6-甲基鸟嘌呤-DNA 甲基转移酶（MGMT）mRNA 表达降低。因此，这些观察结果表明，雷公藤红素诱导 DNA 损伤并抑制 DNA 损伤和修复相关基因表达（mRNA），这可能是雷公藤红素在体外抑制 A375.S2 细胞生长的因素。
