| 徐凯红.白花丹醌对白血病细胞DNA甲基转移酶的作用[J].浙江中西医结合杂志,2021,31(12): |
| 白花丹醌对白血病细胞DNA甲基转移酶的作用 |
| Effect of Plumbagin on DNA methyltransferase in leukemia cells |
| 投稿时间：2021-05-24 修订日期：2021-11-12 |
| DOI： |
| 中文关键词: 白花丹醌 甲基转移酶 白血病细胞 DNA甲基化 |
| 英文关键词:plumbagin methyltransferase leukemia cell DNA methylation |
| 基金项目:宁波市自然科学(2017A610210)(2016A610152)作者单位：1.宁波市第一医院血液科(宁波 316000)；2.宁波市第一医院干细胞实验室(宁波 316000)通讯作者：徐凯红,Tel:0574-87085591;E-mail:xukaho@163.com ,严笑1,杨澍君2,张怡2,牧启田 2,徐志娟1,欧阳桂芳1 |
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| 中文摘要: |
| 目的 探讨中草药提取物白花丹醌对人白血病细胞系HL-60甲基化的影响，并对其相关机制做初步研究。方法MTS法确定不影响细胞生长的白花丹醌研究浓度；白花丹醌研究浓度作用于HL-60细胞不同时间，ELISA法测定其甲基化水平和DNA甲基转移酶(DNMT)活性；实时荧光定量PCR，评估DNMT酶类基因mRNA表达水平。结果 选定白花丹醌0.8μmol/L作为研究浓度，作用18h后，和对照组比较出现甲基化水平显著下降（P<0.05）, 经活性氧（ROS）阻滞剂的预处理，白花丹醌去甲基化作用被明显抑制，使得预处理组与对照组甲基化水平无明显统计学差异[(2.11±0.16)ng 对比(2.27±0.21)ng（P>0.05）]；白花丹醌作用HL-60细胞12h后,出现DNMT酶总体活性的下降[（0.91±0.11）OD/h/μg对比对照组（1.24±0.91）OD/h/μg （P<0.05）]，作用24h后下降更明显（P<0.01）；白花丹醌对DNMT1,DNMT3b基因的表达具有明显抑制作用（P<0.05），对DNMT3a基因表达的作用无统计学意义。结论 低浓度白花丹醌以ROS为重要介质，能够诱导白血病细胞去甲基化，这种作用与抑制DNMT活性，抑制DNMT1和DNMT3b基因的表达有关。 |
| 英文摘要: |
| Objective To investigate the effect of Plumbagin from Chinese Herbon on methylation of human leukemia cell line HL-60.Methods Determining the concentration of Plumbagin for the study by MTS method; Being treated by different time with Plumbagin, the methylation level and the activity of methyltransferase (DNMT) were studied by ELISA; mRNA expression levels of DNMT genes were evaluated by real-time quantitative PCR. Result The 0.8μmol/L of Plumbagin was selected as the study concentration. A decrease in methylation level was observed after 18h-treatment compared with the control group （P<0.05）; Pretreated with the ROS scavenger, the demethylation was greatly inhibited, without significant difference compared with the control group [(2.11±0.16)ng vs. (2.27±0.21)ng（P>0.05）];The total activity of DNMT decreased after treated with Plumbagin for 12h [(0.91±0.11) OD/h/μg vs.control group (1.24±0.91) OD/h/μg（P<0.05）]. The decrease was more obvious after 24h treatment (P<0.01)].Plumbagin could dampen the expression of DNMT1 and DNMT3b genes (P<0.05) ,but without significant role on DNMT3a.Conclusion With ROS as an important mediator, low concentration of Plumbagin can induce demethylation of leukemia cells, partly depending on the reduction of methyltransferase activity and the expression of DNMT1 and DNMT3b. |
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