5-氮杂-2-脱氧胞苷通过去甲基化胸苷磷酸化酶启动子增强 5-氟尿嘧啶的敏感性。
5-Aza-2-deoxycytidine Enhances the Sensitivity of 5-Fluorouracil by Demethylation of the Thymidine Phosphorylase Promoter.
机构信息
Department of Clinical Pharmacy and Pharmacology, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan.
Department of Pharmacy, University of Miyazaki Hospital, Miyazaki, Japan.
出版信息
Anticancer Res. 2019 Aug;39(8):4129-4136. doi: 10.21873/anticanres.13571.
BACKGROUND/AIM: 5-Aza-2-deoxycytidine (5-Aza-CdR) enhances the sensitivity to 5-fluorouracil (5-FU), but the molecular mechanism is not fully understood. The aim of this study was to investigate the molecular mechanism that enhances the sensitivity to 5-FU treated with 5-Aza-CdR via thymidine phosphorylase (TP).
MATERIALS AND METHODS
The sensitivity to drugs was determined on several cancer cell lines by the MTT assay. Protein and mRNA levels were examined by immunoblot and RT-PCR, respectively. Gene silencing, binding of Sp1 to DNA and methylation of DNA was performed by siRNA, ChIP assay and sodium bisulfate genomic sequencing, respectively.
RESULTS
Sp1-binding sites in the TP promoter were methylated in epidermoid carcinoma. 5-Aza-CdR demethylated Sp1-binding sites and enhanced sensitivity to 5-FU.
CONCLUSION
Demethylation of Sp1-binding sites by 5-Aza-CdR was a key factor enhancing 5-FU sensitivity, which may enable more effective treatments for cancer patients with the combination of 5-Aza-CdR and 5-FU.
背景/目的：5-氮杂-2-脱氧胞苷（5-Aza-CdR）可增强 5-氟尿嘧啶（5-FU）的敏感性，但分子机制尚不完全清楚。本研究旨在通过胸苷磷酸化酶（TP）探讨 5-Aza-CdR 增强 5-FU 敏感性的分子机制。
材料和方法
通过 MTT 法在几种癌细胞系中测定药物敏感性。通过免疫印迹和 RT-PCR 分别检测蛋白和 mRNA 水平。通过 siRNA、ChIP 测定和亚硫酸氢钠基因组测序分别进行基因沉默、Sp1 与 DNA 的结合和 DNA 甲基化。
结果
表皮样癌细胞中 TP 启动子中的 Sp1 结合位点发生甲基化。5-Aza-CdR 去甲基化 Sp1 结合位点并增强了对 5-FU 的敏感性。
结论
5-Aza-CdR 对 Sp1 结合位点的去甲基化是增强 5-FU 敏感性的关键因素，这可能使更多癌症患者通过 5-Aza-CdR 和 5-FU 的联合治疗获得更有效的治疗效果。
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