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Circular RNA circRHOT1 promotes hepatocellular carcinoma progression by initiation of NR2F6 expression
BACKGROUND: Increasing evidence has revealed a close relationship between non-coding RNAs and cancer progression. Circular RNAs (circRNAs), a recently identified new member of non-coding RNAs, are demonstrated to participate in diverse biological processes, such as development, homeostatic maintenan...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6639939/ https://www.ncbi.nlm.nih.gov/pubmed/31324186 http://dx.doi.org/10.1186/s12943-019-1046-7 |
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author | Wang, Liyan Long, Haiyan Zheng, Qinghua Bo, Xiaotong Xiao, Xuhua Li, Bin |
author_facet | Wang, Liyan Long, Haiyan Zheng, Qinghua Bo, Xiaotong Xiao, Xuhua Li, Bin |
author_sort | Wang, Liyan |
collection | PubMed |
description | BACKGROUND: Increasing evidence has revealed a close relationship between non-coding RNAs and cancer progression. Circular RNAs (circRNAs), a recently identified new member of non-coding RNAs, are demonstrated to participate in diverse biological processes, such as development, homeostatic maintenance and pathological responses. The functions of circRNAs in cancer have drawn wide attention recently. Until now, the expression patterns and roles of circRNAs in hepatocellular carcinoma (HCC) have remained largely unknown. METHODS: Bioinformatics method was used to screen differentially expressed novel circRNAs in HCC. Northern blotting, qRT-PCR, in situ hybridization (ISH) and RNA-FISH were utilized to analyzed the expression of circRHOT1 in HCC tisues.CCK8, colony formation, EdU assays were used to analyze proliferation of HCC cells. Transwell assay was utilized to analyze HCC cell migration and invasion. FACS was used for apoptosis analysis. Xenograft experiments were used to analyze tumor growth in vivo. Mass spectrum, RNA pulldown, RIP and EMSA was utilized to test the interaction between circRHOT1 and TIP60. RNA-sequencing method was used to analyze the downstream target gene of circRHOT1. RESULTS: We identified circRHOT1 (hsa_circRNA_102034) as a conserved and dramatically upregulated circRNA in HCC tissues. HCC patients displaying high circRHOT1 level possessed poor prognosis. Through in vitro and in vivo experiments, we demonstrated circRHOT1 significantly promoted HCC growth and metastasis. Regarding the mechanism, we conducted a RNA pulldown with a biotin-labeled circRHOT1-specific probe and found that circRHOT1 recruited TIP60 to the NR2F6 promoter and initiated NR2F6 transcription. Moreover, NR2F6 knockout inhibited growth, migration and invasion, whereas rescuing NR2F6 in circRHOT1-knockout HCC cells rescued the proliferation and metastasis abilities of HCC cells. CONCLUSION: Taken together, circRHOT1 inhibits HCC development and progression via recruiting TIP60 to initiate NR2F6 expression, indicating that circRHOT1 and NR2F6 may be potential biomarkers for HCC prognosis. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12943-019-1046-7) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6639939 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-66399392019-07-29 Circular RNA circRHOT1 promotes hepatocellular carcinoma progression by initiation of NR2F6 expression Wang, Liyan Long, Haiyan Zheng, Qinghua Bo, Xiaotong Xiao, Xuhua Li, Bin Mol Cancer Research BACKGROUND: Increasing evidence has revealed a close relationship between non-coding RNAs and cancer progression. Circular RNAs (circRNAs), a recently identified new member of non-coding RNAs, are demonstrated to participate in diverse biological processes, such as development, homeostatic maintenance and pathological responses. The functions of circRNAs in cancer have drawn wide attention recently. Until now, the expression patterns and roles of circRNAs in hepatocellular carcinoma (HCC) have remained largely unknown. METHODS: Bioinformatics method was used to screen differentially expressed novel circRNAs in HCC. Northern blotting, qRT-PCR, in situ hybridization (ISH) and RNA-FISH were utilized to analyzed the expression of circRHOT1 in HCC tisues.CCK8, colony formation, EdU assays were used to analyze proliferation of HCC cells. Transwell assay was utilized to analyze HCC cell migration and invasion. FACS was used for apoptosis analysis. Xenograft experiments were used to analyze tumor growth in vivo. Mass spectrum, RNA pulldown, RIP and EMSA was utilized to test the interaction between circRHOT1 and TIP60. RNA-sequencing method was used to analyze the downstream target gene of circRHOT1. RESULTS: We identified circRHOT1 (hsa_circRNA_102034) as a conserved and dramatically upregulated circRNA in HCC tissues. HCC patients displaying high circRHOT1 level possessed poor prognosis. Through in vitro and in vivo experiments, we demonstrated circRHOT1 significantly promoted HCC growth and metastasis. Regarding the mechanism, we conducted a RNA pulldown with a biotin-labeled circRHOT1-specific probe and found that circRHOT1 recruited TIP60 to the NR2F6 promoter and initiated NR2F6 transcription. Moreover, NR2F6 knockout inhibited growth, migration and invasion, whereas rescuing NR2F6 in circRHOT1-knockout HCC cells rescued the proliferation and metastasis abilities of HCC cells. CONCLUSION: Taken together, circRHOT1 inhibits HCC development and progression via recruiting TIP60 to initiate NR2F6 expression, indicating that circRHOT1 and NR2F6 may be potential biomarkers for HCC prognosis. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12943-019-1046-7) contains supplementary material, which is available to authorized users. BioMed Central 2019-07-19 /pmc/articles/PMC6639939/ /pubmed/31324186 http://dx.doi.org/10.1186/s12943-019-1046-7 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Wang, Liyan Long, Haiyan Zheng, Qinghua Bo, Xiaotong Xiao, Xuhua Li, Bin Circular RNA circRHOT1 promotes hepatocellular carcinoma progression by initiation of NR2F6 expression |
title | Circular RNA circRHOT1 promotes hepatocellular carcinoma progression by initiation of NR2F6 expression |
title_full | Circular RNA circRHOT1 promotes hepatocellular carcinoma progression by initiation of NR2F6 expression |
title_fullStr | Circular RNA circRHOT1 promotes hepatocellular carcinoma progression by initiation of NR2F6 expression |
title_full_unstemmed | Circular RNA circRHOT1 promotes hepatocellular carcinoma progression by initiation of NR2F6 expression |
title_short | Circular RNA circRHOT1 promotes hepatocellular carcinoma progression by initiation of NR2F6 expression |
title_sort | circular rna circrhot1 promotes hepatocellular carcinoma progression by initiation of nr2f6 expression |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6639939/ https://www.ncbi.nlm.nih.gov/pubmed/31324186 http://dx.doi.org/10.1186/s12943-019-1046-7 |
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