Objective:Metastasis is one of the key causes of high mortality in lung cancer.Aberrant DNA methylation is a common event in metastatic lung cancer.We aimed to identify new epigenetic regulation of metastasis-associated genes and characterize their effects on lung cancer progression.Methods:We screened genes associated with non-small cell lung cancer(NSCLC)metastasis by integrating datasets from the Gene Expression Omnibus(GEO)database.We obtained epigenetic-regulated candidate genes by analyzing the expression profile of demethylation genes.By overlapping analysis,epigenetically modulated metastasis-associated genes were obtained.Kaplan-Meier plotter(KM plotter)was utilized to assess the overall survival(OS)of stomatin in lung cancer.Immunohistochemistry(IHC)was conducted to determine the association between stomatin and metastasis-associated clinical indicators.Both in vitro and in vivo assays were performed to investigate the potential role of stomatin in metastasis.The regulation mechanisms of transforming growth factorβ1(TGFβ1)on stomatin were determined by Sequenom MassARRAY quantitative methylation and western blot assays.Results:A series of bioinformatic analyses revealed stomatin as the metastasis-associated gene regulated by DNA methylation.The KM plotter analysis showed a positive association between stomatin and the OS of lung cancer.IHC analysis indicated that the decreased stomatin expression is linked with advanced TNM stage.Loss-and gainof-function experiments displayed that stomatin could inhibit the migration and invasion of NSCLC cells.Furthermore,TGFβ1 repressed stomatin expression during epithelial-to-mesenchymal transition(EMT).The negative correlation between stomatin and TGFβ1 was also validated in advanced stage III lung tumor samples.The underlying mechanism by which TGFβ1 inhibits stomatin is due in part to DNA methylation.Conclusions:Our results suggest that stomatin may be a target for epigenetic regulation and can be used to prevent metastatic diseases.
Huaying AnXiao MaMingyi LiuXiaotong WangXundong WeiWei YuanJie Ma
目的:研究stomatin样蛋白2(stomatin-like protein 2,SLP-2)对人子宫内膜基质细胞系T-HESC的增殖和分化的调控作用。方法:培养子宫内膜基质细胞系T-HESC,体外诱导蜕膜化,RT-q PCR、Western blot和免疫荧光方法检测SLP-2在体外诱导蜕膜化模型中的表达变化以及蜕膜化标志分子催乳素(prolactin,PRL)、胰岛素样生长因子结合蛋白1(insulin like growth factor binding protein 1,IGFBP1)和Desmin的表达。转染特异性SLP-2干扰小RNA(si RNA),敲低SLP-2的表达,分析敲低SLP-2表达对基质细胞增殖和分化(蜕膜化)的影响。结果:体外诱导蜕膜化条件下,诱导蜕膜化标志物IGFBP1和PRL的表达明显增加;Desmin显著表达于在体外诱导蜕膜化的细胞中;SLP-2表达明显升高。在蜕膜化细胞中转染SLP-2 si RNA敲低SLP-2表达,蜕膜化行为受到显著抑制;细胞增殖标志分子CCND3表达明显下降,细胞增殖降低。结论:SLP-2的表达可影响人子宫内膜基质细胞细胞系T-HESC的增殖和分化。
