https://scholars.lib.ntu.edu.tw/handle/123456789/503598
標題: | Mechanisms involved in the activation of C/EBPα by small activating RNA in hepatocellular carcinoma | 作者: | Zhao X. Reebye V. Hitchen P. Fan J. Jiang H. S?trom P. Rossi J. Habib N.A. KAI-WEN HUANG |
公開日期: | 2019 | 出版社: | Nature Publishing Group | 卷: | 38 | 期: | 18 | 起(迄)頁: | 3446-3457 | 來源出版物: | Oncogene | 摘要: | Hepatocellular carcinoma (HCC) is generally accompanied by high mortality and low cure rate. CCAAT enhancer-binding proteins (CEBPs) are transcriptional regulators that play a key role in maintaining liver function. Altered expression of C/EBPα and C/EBPβ occurs in many tumours including HCC. saRNAs are small double-stranded RNAs that enhance target gene expression at the transcriptional level. In this report, we activate CEPBA with saRNAs and suppress CEBPB with siRNAs in cells that represent three different degrees of HCC. We performed functional assays to investigate the effects of enhancing C/EBPα and its downstream targets, p21 and albumin across these lines. We also used Mass-spectrometry (MS) subsequent to a ChIP pull-down assay to characterise the components of the protein complex involved in regulating saRNA function. Putative saRNA interacting protein candidates that were identified by MS were knocked-down with siRNAs to investigate its impact on saRNA activity. We confirmed CEBPA-saRNA decreased proliferation and migration in the differentiated lines (HepG3/Hep3B). The undifferentiated line (PLCPRF5) showed saRNA-induced increase in CEBPA but with no loss in proliferation. This effect was reversed when CEBPB was suppressed with CEBPB-siRNA. When interrogating saRNA mode of action; three saRNA interacting proteins, CTR9, HnRNPA2/B1 and DDX5 were identified by MS. Targeted knock-down of these two proteins (by siRNA) abrogated saRNA activity. This study provides insight into how different HCC lines are affected by CEBPA-saRNAs and that endogenous abundance of CEBPB and saRNA accessory proteins may dictate efficacy of CEBPA-saRNA when used in a therapeutic context. ? 2019, Springer Nature Limited. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85059962081&doi=10.1038%2fs41388-018-0665-6&partnerID=40&md5=e9ecf6e3a5dabf3338132a8f95c83707 https://scholars.lib.ntu.edu.tw/handle/123456789/503598 |
ISSN: | 0950-9232 | DOI: | 10.1038/s41388-018-0665-6 | SDG/關鍵字: | albumin; CCAAT enhancer binding protein alpha; CCAAT enhancer binding protein beta; CTR9 protein; DEAD box protein; DEAD box protein 5; heterogeneous nuclear ribonucleoprotein; heterogeneous nuclear ribonucleoprotein A2; oligonucleotide; protein p21; RNA; small activating RNA; small interfering RNA; transcription factor; unclassified drug; CCAAT enhancer binding protein; CEBPA protein, human; DEAD box protein; heterogeneous nuclear ribonucleoprotein group A B; nuclear protein; Article; cell migration; cell proliferation; chromatin immunoprecipitation; controlled study; downstream processing; gene function; gene knockdown; hepatocellular carcinoma cell line; liver cell carcinoma; mass spectrometry; priority journal; protein analysis; protein expression; transcription initiation; cell differentiation; gene expression regulation; genetics; Hep-G2 cell line; human; liver cell carcinoma; liver tumor; tumor cell line; Carcinoma, Hepatocellular; CCAAT-Enhancer-Binding Proteins; Cell Differentiation; Cell Line, Tumor; DEAD-box RNA Helicases; Gene Expression Regulation; Hep G2 Cells; Heterogeneous-Nuclear Ribonucleoprotein Group A-B; Humans; Liver Neoplasms; Nuclear Proteins; RNA, Small Interfering |
顯示於: | 醫學系 |
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