中国肿瘤生物治疗杂志2017，Vol.24Issue(11)：1254-1259,6.DOI:10.3872/j.issn.1007-385x.2017.11.002
桥接整合因子1去甲基化诱导细胞周期阻滞抑制食管鳞状细胞癌EC109细胞增殖
Demethylation of bridging integrator-1 inhibits proliferation of the esophageal squamous cell carcinoma EC109 cell through inducing cell cycle arrest
摘要
Abstract
Objective:To analyze effect of demethylation of bridging integrator-1 (Bin1) on expression of Bin1 gene and proliferation ability of the esophageal squamous cell carcinoma (ESCC) EC109 cell,and preliminarily to explore its possible action mechanism.Methods:Methylation specific Polymerase Chain Reaction (MSP) assay was used to detect methylation status of Bin1 promoter region in the EC 109 cell after treatment with demethylation drug,5-Aza-2'-deoxycytidine (5-Aza-dc).qPCR,Western blotting and MTT assays were used to detect effect of treatment with 5-Aza-dc only and treatment with 5-Aza-dc plus transfection with Bin1 gene interference fragments (Bin1 siRNA) on expressions of Bin1 mRNA and its protein in the EC109 cell as well as proliferation ability of the EC109 cell respectively.Cell cycles of the EC109 cell and expressions of cell cycle-related proteins (Cyclin D1 and CDK4) in the EC109 cell were tested by flow cytometry assay and Western blotting respectively.Results:Demethylation of Bin1 gene promoter region occurred in the EC109 cell after treatment with 5-Aza-dc.In the EC 109 cell treated with 5-Aza-dc,expressions of Bin1 mRNA and its protein obviously up-regulated,proliferation ability of the EC109 cell remarkably decreased,the EC109 cell was blocked at G0/G1 stage,proportion of the EC109 cell at S stage evidently reduced,as well as expressions of Cyclin D1 and CDK4 proteins were markedly down-regulated (all P＜0.05).After the EC109 cell in demethylation status was transfected with Bin1 siRNA,expressions of Bin1 mRNA and its protein significantly decreased and proliferation ability of the EC109 cell evidently enhanced (all P＜0.05).Conclusion:Bin1 gene promoter region in the EC109 cell appeared in complete demethylation status.Demethylation of 5-Aza-dc could enhance expression of Bin1 in the EC109 cell of ESCC,which could induce cell cycle arrest and inhibit proliferation of the EC 109 cell through decrease of cell cycle-related protein expression.It was confirmed that epigenetic changes could be related to malignant proliferation of the ESCC cell,which could provide novel idea for therapy of the esophageal carcinoma.关键词
食管鳞状细胞癌/EC109细胞/甲基化/桥接整合因子1基因/细胞增殖/细胞周期Key words
esophageal squamous cell carcinoma (ESCC)/EC109 cell/methylation/bridging integrator-1 gene (Bin1)/cell proliferation/cell cycle分类
医药卫生引用本文复制引用
刘天旭,张翔宇,邓佳,王雪晓,王佳丽,刘丽华..桥接整合因子1去甲基化诱导细胞周期阻滞抑制食管鳞状细胞癌EC109细胞增殖[J].中国肿瘤生物治疗杂志,2017,24(11):1254-1259,6.基金项目
国家自然科学基金资助项目(No.81201607) （No.81201607）
河北省杰出青年基金资助项目(No.H2014206320) （No.H2014206320）
河北省高层次人才培养项目资助(No.A201401040). Project supported by the National Natural Science Foundation of China (No.81201607),the Outstanding Youth Foundation of Hebei Province (H2014206320),and the Foundation for High Level Talents of Hebei Province (No.A201401040) （No.A201401040）
