HaCaT细胞中FUT4表达与其启动子区甲基化的相关性分析

doi:10.16288/j.yczz.2015.01.007

遗传 ›› 2015, Vol. 37 ›› Issue (1): 48-54.doi: 10.16288/j.yczz.2015.01.007

HaCaT细胞中FUT4表达与其启动子区甲基化的相关性分析

李洪艳¹, 佟少明¹, 燕秋²

1. 辽宁师范大学生命科学学院,大连 116081;
2. 大连医科大学基础医学院,生物化学与分子生物学教研室,大连 116044

收稿日期:2014-07-09 修回日期:2014-08-19 出版日期:2015-01-20 发布日期:2015-01-20
通讯作者: 燕秋,博士,教授,研究方向：糖生物学。E-mail: yanqiu63@126.com E-mail:yanqiu63@126.com
作者简介:李洪艳,博士,副教授,研究方向：肿瘤糖生物学,表观遗传学。Tel: 0411-85827090;E-mail: L-hongyan@163.com

Correlation between FUT4 expression and its promoter methylation in HaCaT cells

Hongyan Li¹, Shaoming Tong¹, Qiu Yan²

1. College of Life Science, Liaoning Normal University, Dalian 116081, China; 2. Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian 116044, China

Received:2014-07-09 Revised:2014-08-19 Online:2015-01-20 Published:2015-01-20

摘要/Abstract

摘要： 岩藻糖基转移酶Ⅳ(Fucosyltransferase Ⅳ,FUT4)在正常细胞中表达量很低,但其低表达的调控机制以及是否受其启动子甲基化调控并不十分清楚。文章采用Western blot、免疫荧光和Real-time PCR的方法检测正常人永生化表皮细胞系HaCaT细胞FUT4的表达,观察DNA甲基转移酶抑制剂5-aza-dC处理对FUT4表达的影响。应用甲基化特异性PCR方法分析HaCaT细胞中FUT4启动子甲基化状态。结果表明,HaCaT细胞中FUT4的表达水平明显低于人表皮鳞癌细胞A431和SCC12。5 μmol/L的5-aza-dC处理72 h的HaCaT细胞,其FUT4 mRNA水平明显升高,并且与未经5-aza-dC处理的对照组相比,U引物扩增检测到的产物量增加,M 引物扩增检测到的产物量明显减少。这些结果表明,HaCaT细胞中FUT4的低表达可能与其启动子区CpG岛甲基化有关。

关键词: 岩藻糖基转移酶Ⅳ, 低表达, 甲基化, HaCaT细胞

Abstract: The expression level of fucosyltransferase Ⅳ (FUT4) is low in normal cells. The mechanism underlying regulation of FUT4 expression in normal cells remains elusive. In this study, Western blot, immunofluorescence and real-time PCR were used to analyze FUT4 expression in the immortalized human keratinocytes cells HaCaT. Methylated-specific PCR was used to investigate methylation status of FUT4 promoter. The results showed that the FUT4 expression level was significantly lower in HaCaT cells than squamous carcinoma cells A431 and SCC12. FUT4 mRNA expression was increased in HaCaT cells treated by 5-aza-dC (5 μmol/L), an inhibitor of DNA methyltransferase. Furthermore, using the primers to amplify the methylated fragment yielded PCR products and no products were yielded by the primers to amplify the unmethylated fragment in HaCaT cells. Unmethylated PCR products were obtained in HaCaT cells treated by 5-aza-dC, while methylated PCR products were not detected. These results suggest that the lower expression of FUT4 in HaCaT cells may be correlated with the methylation of CpG island in FUT4 promoter.

Key words: FUT4, low expression, DNA methylation, HaCaT cells

李洪艳, 佟少明, 燕秋. HaCaT细胞中FUT4表达与其启动子区甲基化的相关性分析[J]. 遗传, 2015, 37(1): 48-54.

Hongyan Li, Shaoming Tong, Qiu Yan. Correlation between FUT4 expression and its promoter methylation in HaCaT cells[J]. HEREDITAS(Beijing), 2015, 37(1): 48-54.

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HaCaT细胞中FUT4表达与其启动子区甲基化的相关性分析

Correlation between FUT4 expression and its promoter methylation in HaCaT cells

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