Identification of downstream metastasis-associated target genes regulated by LSD1 in colon cancer cells.
Identification of downstream metastasis-associated target genes regulated by LSD1 in colon cancer cells.
复制标题
结肠癌细胞中LSD1调控的下游转移相关靶基因的鉴定
DOI：
10.18632/oncotarget.14778
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发表时间：
2017-03-21
期刊：
影响因子：
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通讯作者：
Du J
中科院分区：
文献类型：
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作者：
Chen J;Ding J;Wang Z;Zhu J;Wang X;Du J
目的筛选结肠癌细胞中赖氨酸特异性脱甲基酶1（LSD 1）调控的下游靶基因，探讨LSD 1影响结肠癌侵袭转移的分子机制。方法通过基因表达谱分析，获得两种人结肠癌细胞系中小干扰RNA-LSD 1和LSD 1过表达调控的下游靶基因的表达变化。使用Affyscore Human Transcriptome Array 2.0鉴定差异表达基因（DEG）。通过Gene Ontology和Pathway Studio从DEG中筛选出与增殖、转移和侵袭相关的LSD 1靶基因。随后，四个关键基因（CABYR，FOXF 2，TLE 4和CDH 1）被计算预测为转移相关的LSD 1靶基因。在RT-PCR和Western blot验证后，应用ChIp-PCR检测LSD 1-靶基因启动子结合的LSD 1的占有率。结果LSD 1基因沉默的SW 620细胞中，共有3633个DEGs表达上调，4642个DEGs表达下调。在LSD 1过表达的HT-29细胞中，分别有4047个DEG和4240个DEG被上调和下调。RT-PCR和Western blot验证了芯片分析结果。ChIP实验结果表明，LSD 1可能是靶基因CABYR和CDH 1的负调控因子。LSD 1的表达水平与LSD 1靶基因启动子区组蛋白H3赖氨酸4（H3 K4）的单甲基化和二甲基化呈负相关。在CABYR和CDH 1的启动子区域没有检测到H3赖氨酸9甲基化的显著单甲基化和二甲基化。结论LSD 1缺失可能通过增强靶基因启动子H3 K4的二甲基化作用而促进CABYR和CDH 1表达。LSD 1过表达通过降低LSD 1靶基因启动子H3 K4的单甲基化和二甲基化水平，介导CABYR和CDH 1表达下调。CABYR和CDH 1可能是LSD 1在结肠癌发生中的潜在靶基因。
Purpose This study aims to identify downstream target genes regulated by lysine-specific demethylase 1 (LSD1) in colon cancer cells and investigate the molecular mechanisms of LSD1 influencing invasion and metastasis of colon cancer. Method We obtained the expression changes of downstream target genes regulated by small-interfering RNA-LSD1 and LSD1-overexpression via gene expression profiling in two human colon cancer cell lines. An Affymetrix Human Transcriptome Array 2.0 was used to identify differentially expressed genes (DEGs). We screened out LSD1-target gene associated with proliferation, metastasis, and invasion from DEGs via Gene Ontology and Pathway Studio. Subsequently, four key genes (CABYR, FOXF2, TLE4, and CDH1) were computationally predicted as metastasis-related LSD1-target genes. ChIp-PCR was applied after RT-PCR and Western blot validations to detect the occupancy of LSD1-target gene promoter-bound LSD1. Result A total of 3633 DEGs were significantly upregulated, and 4642 DEGs were downregulated in LSD1-silenced SW620 cells. A total of 4047 DEGs and 4240 DEGs were upregulated and downregulated in LSD1-overexpressed HT-29 cells, respectively. RT-PCR and Western blot validated the microarray analysis results. ChIP assay results demonstrated that LSD1 might be negative regulators for target genes CABYR and CDH1. The expression level of LSD1 is negatively correlated with mono- and dimethylation of histone H3 lysine4(H3K4) at LSD1- target gene promoter region. No significant mono-methylation and dimethylation of H3 lysine9 methylation was detected at the promoter region of CABYR and CDH1. Conclusion LSD1- depletion contributed to the upregulation of CABYR and CDH1 through enhancing the dimethylation of H3K4 at the LSD1-target genes promoter. LSD1- overexpression mediated the downregulation of CABYR and CDH1expression through decreasing the mono- and dimethylation of H3K4 at LSD1-target gene promoter in colon cancer cells. CABYR and CDH1 might be potential LSD1-target genes in colon carcinogenesis.