論文 - 詳細
| RRC ID | 44575 |
|---|---|
| 著者 | Iwaki J, Kikuchi K, Mizuguchi Y, Kawahigashi Y, Yoshida H, Uchida E, Takizawa T. |
| タイトル | MiR-376c down-regulation accelerates EGF-dependent migration by targeting GRB2 in the HuCCT1 human intrahepatic cholangiocarcinoma cell line. |
| ジャーナル | PLoS One |
| Abstract |
MicroRNA miR-376c was expressed in normal intrahepatic biliary epithelial cells (HIBEpiC), but was significantly suppressed in the HuCCT1 intrahepatic cholangiocarcinoma (ICC) cell line. The biological significance of the down-regulation of miR-376c in HuCCT1 cells is unknown. We hypothesized that miR-376c could function as a tumor suppressor in these cells. To test this hypothesis, we sought the targets of miR-376c, and characterized the effect of its down-regulation on HuCCT1 cells. We performed proteomic analysis of miR-376c-overexpressing HuCCT1 cells to identify candidate targets of miR-376c, and validated these targets by 3'-UTR reporter assay. Transwell migration assays were performed to study the migratory response of HuCCT1 cells to miR-376c overexpression. Furthermore, microarrays were used to identify the signaling that were potentially involved in the miR-376c-modulated migration of HuCCT1. Finally, we assessed epigenetic changes within the potential promoter region of the miR-376c gene in these cells. Proteomic analysis and subsequent validation assays showed that growth factor receptor-bound protein 2 (GRB2) was a direct target of miR-376c. The transwell migration assay revealed that miR-376c significantly reduced epidermal growth factor (EGF)-dependent cell migration in HuCCT1 cells. DNA microarray and subsequent pathway analysis showed that interleukin 1 beta and matrix metallopeptidase 9 were possible participants in EGF-dependent migration of HuCCT1 cells. Bisulfite sequencing showed higher methylation levels of CpG sites upstream of the miR-376c gene in HuCCT1 relative to HIBEpiC cells. Combined treatment with the DNA-demethylating agent 5-aza-2'-deoxycytidine and the histone deacetylase inhibitor trichostatin A significantly upregulated the expression of miR-376c in HuCCT1 cells. We revealed that epigenetic repression of miR-376c accelerated EGF-dependent cell migration through its target GRB2 in HuCCT1 cells. These findings suggest that miR-376c functions as a tumor suppressor. Since metastasis is the major cause of death in ICC, microRNA manipulation could lead to the development of novel anti-cancer therapy strategies for ICC. |
| 巻・号 | 8(7) |
| ページ | e69496 |
| 公開日 | 2013-1-1 |
| DOI | 10.1371/journal.pone.0069496 |
| PII | PONE-D-13-09943 |
| PMID | 23922722 |
| PMC | PMC3724868 |
| MeSH | Bile Duct Neoplasms / genetics Bile Duct Neoplasms / pathology Bile Ducts, Intrahepatic Cell Line, Tumor Cell Movement / drug effects Cell Movement / genetics* Cholangiocarcinoma / genetics* Cholangiocarcinoma / pathology* DNA Methylation / drug effects DNA Methylation / genetics Down-Regulation / drug effects Down-Regulation / genetics* Epidermal Growth Factor / pharmacology* Epigenesis, Genetic / drug effects GRB2 Adaptor Protein / metabolism* Gene Expression Regulation, Neoplastic / drug effects Gene Regulatory Networks / drug effects Gene Regulatory Networks / genetics Humans Liver Neoplasms / genetics* Liver Neoplasms / pathology* MicroRNAs / genetics* MicroRNAs / metabolism Neoplasm Proteins / metabolism Proteomics Reproducibility of Results |
| IF | 2.74 |
| 引用数 | 41 |
| WOS 分野 | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 3 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | TKKK(RCB1907) HuH-28(RCB1943) TFK-1(RCB2537) |