[1]白倩,谢琦,彭晓莉,等.二氢杨梅素通过抑制甲基转移酶诱导人乳腺癌MCF-7细胞PTEN基因去甲基化[J].第三军医大学学报,2014,36(01):20-24.
 Bai Qian,Xie Qi,Peng Xiaoli,et al.Dihydromyricetin induces PTEN demethylation by down-regulating DNA methyltransferases in human MCF-7 breast cancer cells[J].J Third Mil Med Univ,2014,36(01):20-24.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
36卷
期数:
2014年第01期
页码:
20-24
栏目:
论著
出版日期:
2014-01-15

文章信息/Info

Title:
Dihydromyricetin induces PTEN demethylation by down-regulating DNA methyltransferases in human MCF-7 breast cancer cells
作者:
白倩谢琦彭晓莉常徽朱俊东糜漫天
第三军医大学军事预防医学院营养与食品安全研究中心,重庆市营养与食品安全重点实验室
Author(s):
Bai Qian Xie Qi Peng Xiaoli Chang Hui Zhu Jundong Mi Mantian
Center for Nutrition and Food Safety, Chongqing Key Labortary of Nutrition and Food Safety, College of Military Preventive Medicine, Third Military Medical University, Chongqing, 400038, China
关键词:
二氢杨梅素乳腺癌细胞PTEN去甲基化DNA甲基转移酶
Keywords:
dihydromyricetin breast cancer cells phosphatase and tensin homology deleted on chromosome ten demethylation DNA methyltransferase
分类号:
R282.71;R73-361;R737.9
文献标志码:
A
摘要:
目的      观察二氢杨梅素对人乳腺癌MCF-7细胞中第10号染色体缺失的磷酸酶和张力蛋白同源基因(phosphatase and tensin homology deleted on chromosome ten,PTEN)甲基化及其表达的影响,并探讨相关作用机制。      方法        以CCK-8法检测细胞活力,qRT-PCR检测PTEN和DNA甲基转移酶(DNA methyltransferase,DNMT)DNMT1、DNMT3a、DNMT3b 的mRNA表达,Western blot法检测PTEN的蛋白表达,荧光法检测细胞DNA甲基转移酶总活性,甲基化特异性PCR法(methylation-specific PCR,MSP)检测PTEN基因甲基化水平。      结果        细胞活力检测结果表明,二氢杨梅素可剂量依赖性降低乳腺癌MCF-7细胞活力(P<0.05);qRT-PCR和Western blot检测结果表明,二氢杨梅素可显著上调PTEN的表达(P<0.05),并呈现剂量-效应关系;MSP检测结果显示,二氢杨梅素可显著诱导MCF-7细胞PTEN基因去甲基化;qRT-PCR和荧光法检测结果显示,二氢杨梅素可显著降低MCF-7细胞DNMT1表达和DNMT活性(P<0.05)。      结论        二氢杨梅素显著诱导乳腺癌细胞PTEN基因去甲基化,促进其表达,其机制可能主要涉及对DNMT1表达和活性的抑制。
Abstract:
Objective        To determine the effects of dihydromyricetin on methylation status and expression of phosphatase and tensin homology deleted on chromosome ten (PTEN) gene in human MCF-7 breast cancer cells and explore the relevant mechanisms.       Methods        Cells viability was assessed by CCK-8 assay in MCF-7 cells after the treatment of dihydromyricetin at 10, 20, 40 and 80 μmol/L for 48 h. The expression of DNA methyltransferases (DNMT, including DNMT1, DNMT3a, and DNMT3b) and PTEN was measured by quantitative real-time PCR and Western blotting. DNMT activity was detected by DNMT activity/inhibition assay kit (fluorometrics). Modified methylation specific PCR (MSP) was used to screen the methylation level of PTEN.       Results        Dihydromyricetin significantly decreased the viability of MCF-7 cells in a dose-dependent manner. The qRT-PCR analysis and Western blotting showed that dihydromyricetin dose-dependently increased the expression of PTEN in MCF-7 cells. MSP showed that dihydromyricetin markedly induced PTEN demethylation in MCF-7 cells. Further studies indicated that dihydromyricetin decreased the expression of DNMT1 and inhibited the activity of DNMT.       Conclusion        Dihydromyricetin significantly induces PTEN demethylation and promotes its expression in breast cancer MCF-7 cells, and this effect may be mainly correlated with inhibition of DNMT1 expression and activity.

参考文献/References:

白倩, 谢琦, 彭晓莉, 等. 二氢杨梅素通过抑制甲基转移酶诱导人乳腺癌MCF-7细胞PTEN基因去甲基化[J].第三军医大学学报,2014,36(1):20-24.

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更新日期/Last Update: 2014-01-06