机构：[a]Soochow University Laboratory of Cancer Molecular Genetics, Medical College of Soochow University, Suzhou 215123, China[b]Department of Thoracic and Cardiovascular Surgery, The Second Affiliated Hospital of Soochow University, Medical College of Soochow University, Suzhou 215004, China[c]Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Soochow University, Medical College of Soochow University, Suzhou 215006, China[d]Department of Radiology, The First Affiliated Hospital of Soochow University, Medical College of Soochow University, Suzhou 215006, China
Objectives: RNF111/Arkadia is a critical regulator of TGF-beta signaling, being required for SMAD3-mediated responses such as TGF-beta-induced repression of E-cadherin. Previous studies show that mutations in RNF111 in human cancers are rare and RNF111 promotes lung tumor metastasis. However, the epigenetic mechanisms underlying the role of RNF111 in non-small cell lung cancer (NSCLC) metastasis remain unknown. Here, we mainly focused on low- (95C) and high-metastatic (95D) NSCLC cell lines, which share a similar genetic background, and investigated the methylation-based regulation of RNF111 expression. Materials and methods: Clonal bisulfite sequencing, real-time qRT-PCR, western blot analysis, luciferase reporter assays, RNA interference, chromatin immunoprecipitation (ChIP) assay and transwell migration and invasion assays were performed on human NSCLC cell lines 95C and 95D. Results: RNF111 was high-expressed in 95D cells, which showed low-level methylation at -459CpG site in RNF111 promoter. The opposite results were obtained in 95C cells. Cell-based and biochemical assays revealed that -459CpG methylation can inhibit RNF111 transcriptional expression by interfering with the recruitment of Sp1 to RNF111 promoter. On TGF-beta stimulation, siRNA-mediated RNF111 knockdown inhibited TGF-beta/Smad signaling activity and Snail (an inducer of metastasis) expression, and enhanced E-cadherin (an epithelial-to-mesenchymal transition marker) expression in 95C and 95D cells. Furthermore, demethylation-induced upregulation of RNF111 enhanced phosphorylation of SMAD3 and Snail expression, and repressed E-cadherin expression in 95C cells expressing low RNF111. Conclusions: Our results suggest that -459CpG methylation in Sp1-binding site of RNF111 promoter transcriptionally decreases RNF111 expression, which inhibits TGF-beta/Smad signaling associated invasion in NSCLC cells. (C) 2015 Elsevier Ireland Ltd. All rights reserved.
第一作者机构：[a]Soochow University Laboratory of Cancer Molecular Genetics, Medical College of Soochow University, Suzhou 215123, China
共同第一作者：
通讯作者：
通讯机构：[b]Department of Thoracic and Cardiovascular Surgery, The Second Affiliated Hospital of Soochow University, Medical College of Soochow University, Suzhou 215004, China[*1]Soochow University Laboratory of Cancer Molecular Genetics, Medical College of Soochow University, 199 Ren’ai Road, Sino-Singapore Industrial Park, Suzhou 215123, China.
推荐引用方式(GB/T 7714)：
Hongbing Chen,Tianjie Yang,Zhe Lei,et al.RNF111/Arkadia is regulated by DNA methylation and affects TGF-beta/Smad signaling associated invasion in NSCLC cells[J].LUNG CANCER.2015,90(1):32-40.doi:10.1016/j.lungcan.2015.07.010.
APA：
Hongbing Chen,Tianjie Yang,Zhe Lei,Longqiang Wang,Haiping Yang...&Hong-Tao Zhang.(2015).RNF111/Arkadia is regulated by DNA methylation and affects TGF-beta/Smad signaling associated invasion in NSCLC cells.LUNG CANCER,90,(1)
MLA：
Hongbing Chen,et al."RNF111/Arkadia is regulated by DNA methylation and affects TGF-beta/Smad signaling associated invasion in NSCLC cells".LUNG CANCER 90..1(2015):32-40
