Tanshinone IIA triggers p53 responses and apoptosis by RNA polymerase II upon DNA minor groove binding
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- 作者：Zhichao Zhang ; Jin Gao ; Yuanyuan Wang ; Ting Song ; Jing Zhang ; Guiye Wu ; Tiantai Zhang ; Guanhua Du
- 关键词：Tanshinone IIA ; DNA minor groove binder ; RNA polymerase II ; Apoptosis
- 刊名：Biochemical Pharmacology
- 出版年：2009
- 出版时间：15 November 2009
- 年：2009
- 卷：78
- 期：10
- 页码：1316-1322
- 全文大小：374 K
文摘
Our previous work has shown that tanshinone IIA (Tan IIA) is a DNA minor groove binder instead of an intercalator as previously thought. In this study, we have further demonstrated that the molecular antitumor pharmacology of Tan IIA is dependent on its groove-binding capability. First, we investigated the structure damage to duplex DNA upon Tan IIA binding using circular dichroism spectra. Subsequently, we performed western blot, flow cytometry analysis, chromatin immunoprecipitation, and quantitative real-time PCR to illustrate the RNAPII degradation, phosphorylation, and distribution along the transcribed gene in H22 cells exposed to Tan IIA. In addition, p53 activation and apoptosis induction in both cultured H22 cells and in mice bearing the ascitic-type H22 were measured following Tan IIA treatment. It was revealed that Tan IIA decreases the level of RNAPII by altering DNA structure. At the low dose range (0.2–4 μM) of Tan IIA exposure, the DNA structure damage results in the inhibition of RNAPII binding to DNA and the initiation of RNAPII phosphorylation, while higher concentrations of Tan IIA (4–20 μM) cause complete phosphorylation and degradation of RNAPII followed by p53 activation and apoptosis. A similar apoptosis induction by RNAPII was observed in animals. Apoptosis of tumor cells from ascitic fluid was not detected until RNAPII levels were downregulated by Tan IIA, which requires 40 mg/kg body weight of Tan IIA. It was concluded that DNA-conformational-damage-dependent RNAPII response upon groove binding is the molecular basis of the antitumor property of Tan IIA, in vivo and in vitro.