引用本文:丁 璇1,李俊枫2,罗子国1,许 舸1,唐 溧1.双氢青蒿素对前列腺癌PC-3细胞UCHL1基因转录调节的影响及机制[J].重庆医科大学学报,2014,38(6):771~775
双氢青蒿素对前列腺癌PC-3细胞UCHL1基因转录调节的影响及机制
Effects of dihydroartemisinin on modulating UCHL1 gene expression in human prostate cancer cell line PC-3 and its mechanism
DOI:
中文关键词:  双氢青蒿素  前列腺癌  泛素羧基末端水解酶1
英文关键词:dihydroartemisinin  prostate cancer  ubiquitin carboxyl-terminal hydrolase L1
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作者单位
丁 璇1,李俊枫2,罗子国1,许 舸1,唐 溧1 1. 重庆医科大学生命科学研究院重庆 4000162. 重庆市垫江县人民医院肿瘤科重庆 408300 
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中文摘要:
      目的:探讨双氢青蒿素(dihydroartemisinin,DHA)对前列腺癌PC-3细胞泛素羧基末端水解酶1(ubiquitin carboxyl-ter-minal hydrolase L1,UCHL1)基因转录调节的影响及机制。方法:将PC-3细胞分为空白组、DHA组及5-杂氮-2’脱氧胞苷(5-Aza-2’-deoxycytidine,5-Aza-CdR)组(2.5 μmol/L 5-Aza-CdR)。空白组为不加药物;DHA组根据浓度的不同再分为DHA1、DHA2、DHA3、DHA4组,DHA终浓度分别为12.5、25、50、100 μmol/L。培养48 h后,收集细胞。应用real-time PCR检测 UCHL1及 DNA甲基化转移酶(DNA methyltransferase,DNMTs)的mRNA表达;亚硫酸氢盐基因组测序法(bisulfite genomic sequencing,BGS)检测UCHL1 CpG岛的甲基化状态;Western blot法检测 UCHL1蛋白的表达。结果:DHA和5-Aza-CdR分别处理细胞后,UCHL1的启动子甲基化水平明显降低(χ2=150.1,P=0.000; χ2=127.7,P=0.000);UCHL1 mRNA表达水平明显上调(P均<0.05);UCHL1蛋白表达水平上升(P均 <0.05),双氢青蒿素处理后甲基化转移酶的 mRNA表达减少(P均<0.05)。结论:DHA可逆转 PC-3细胞 UCHL1基因的甲基化状态,其可能通过下调DNMTs的表达而影响 DNA的甲基化过程。
英文摘要:
      Objective:To investigate the effects of dihydroartemisinin(DHA) on regulating ubiquitin carboxyl-terminal hydrolase L1(UCHL1) gene expression in human prostate cancer cell line PC-3 and its mechanism. Methods:PC-3 cells were divided into six groups including four groups treated with DHA at concentrations of 12.5,25,50,100 μmol/L;5-Aza-CdR group cultured in the media with 5-Aza-2’-deoxycytidine at concentrations of 2.5 μmol/L and blank group cultured with blank media. CpG island of ubiqui-tin carboxyl-terminal hydrolase L1(UCHL1) methylation levels were determined by bisulfite genomic sequencing. UCHL1 mRNA expression levels were determined via realtime fluorescence quantitative PCR whereas the expression levels of UCHL1 protein were determined using Western blot. Results:In the DHA groups and 5-Aza-CdR group,UCHL1 promoter methylation levels were decreased dramatically(χ2=150.1,P=0.000; χ2=127.7,P=0.000). mRNA expression levels of UCHL1 were dramatically increased( χ2=150.1,P=0.000; χ2=127.7,P=0.000). Protein expression levels of UCHL1 were significantly increased(P<0.05). In DHA groups,mRNA ex-pression levels of DNA methyltransferase were decreased(P<0.05). Conclusion:DHA can induce the demethylation of CpG island of UCHL1 through downregulating the expression of DNA methyltransferase in PC-3 cells.
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