乏氧激活人口腔鳞癌细胞DKKL1的表达  

Activation of DKKL1 expression by hypoxia in human OSCC cell lines

作　　者：王建秋[1] 范如意[1] 谢其洋 闫风琴[2] 

机构地区：[1]杭州师范大学医学院生物化学与分子生物学教研室,浙江杭州310036 [2]浙江省肿瘤医院放疗科十九病区

出　　处：《中国老年学杂志》2016年第11期2569-2571,共3页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(No.81301850);浙江省教育厅科研项目(No.Y201328812)

摘　　要：目的探讨常氧和乏氧条件下人口腔鳞癌细胞Dickkopf样蛋白-1(DKKL1)的表达及可能的表观遗传学机制。方法取对数生长期人口腔鳞癌SAS细胞和OECM-1细胞,常氧或乏氧(1.0%O2)条件下培养18 h,采用实时荧光定量PCR方法检测DKKL1 mRNA水平的表达,采用Western印迹方法检测其蛋白水平的表达;进一步采用染色质免疫沉淀(Ch IP)方法检测DKKL1基因启动子上H3K4乙酰化(H3K4ac)和二甲基化(H3K4me2)水平的变化。结果与常氧条件下相比,乏氧使DKKL1在SAS和OECM-1两种口腔鳞癌细胞中的表达均显著升高,其mRNA水平分别为常氧条件下的(4.0±0.4)倍(P<0.01)和(2.4±0.1)倍(P<0.01);Ch IP结果显示,乏氧增加了DKKL1基因启动子上H3K4ac水平,降低了H3K4me2水平。结论乏氧可激活口腔鳞癌细胞DKKL1的表达,该作用可能与乏氧调节DKKL1基因启动子上组蛋白H3K4的乙酰化和二甲基化修饰有关。Objective To investigate the effects of tumor hypoxia microenvironment on expression of DKKL1 in oral squamous cell carcinoma（ OSCC） cells lines and explore its potential epigenetic mechanism.Methods Two kinds of human OSCC cell lines in exponential growth phase,SAS and OECM-1 cells,were cultured in normoxia or hypoxia（ 1. 0% oxygen） conditions for 18 h. Both Real-time PCR and Western blot were applied to detect the change of DKKL1 expression. Chromatin Immunoprecipitation（ Ch IP） experiments were performed to detect the changes of histone modification of acetylated or dimethyl H3K4（ H3K4 ac / H3K4me2） on DKKL1 gene promoter. Results Compared to normoxic conditions,hypoxia（ 1.0%） significantly increased the expression of DKKL1 in both SAS and OECM-1 cells,and the relative mRNA level was（ 4.0±0.4） folds and（ 2.4±0.1） folds of that under normoxia in SAS（ P0.01） or OECM-1（ P 0.01） cells,respectively. Ch IP experiments showed that level of H3K4 ac on promoter of DKKL1 gene was increased,while the level of H3K4me2 was reduced significantly.Conclusions Hypoxia activates the expression of DKKL1,which may be related with the regulation of hypoxia on histone modification of H3K4 ac and H3K4me2 on the promoter of DKKL1 gene.

关 键 词：口腔鳞癌 乏氧 Dickkopf样蛋白-1 组蛋白修饰 

分 类 号：R739.8[医药卫生—肿瘤]