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DANCR promotes HCC progression and regulates EMT by sponging miR‐27a‐3p via ROCK1/LIMK1/COFILIN1 pathway
OBJECTIVES: This research aims to verify that the long non‐coding RNA differentiation antagonizing nonprotein coding RNA (LncRNA DANCR) could modulate the proliferation and metastasis of hepatocellular carcinoma (HCC), and it thus may work as a novel biomarker to render new orientation for early dia...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6668976/ https://www.ncbi.nlm.nih.gov/pubmed/31038266 http://dx.doi.org/10.1111/cpr.12628 |
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author | Guo, Dan Li, Yarui Chen, Yifei Zhang, Dan Wang, Xin Lu, Guifang Ren, Mudan Lu, Xinlan He, Shuixiang |
author_facet | Guo, Dan Li, Yarui Chen, Yifei Zhang, Dan Wang, Xin Lu, Guifang Ren, Mudan Lu, Xinlan He, Shuixiang |
author_sort | Guo, Dan |
collection | PubMed |
description | OBJECTIVES: This research aims to verify that the long non‐coding RNA differentiation antagonizing nonprotein coding RNA (LncRNA DANCR) could modulate the proliferation and metastasis of hepatocellular carcinoma (HCC), and it thus may work as a novel biomarker to render new orientation for early diagnosis and clinical therapy of HCC. MATERIALS AND METHODS: Firstly, qRT‐PCR was used to detect the expression of genes including LncRNA DANCR and miR‐27a‐3p. Next, MTT assay, Ethynyldeoxyuridine (EdU) analysis and clone formation assay were used for investigating cell growth and proliferation. Meanwhile, transwell assay and wound healing assay were applied to evaluate the capacity of cell metastasis and motility, respectively. In addition, bioinformatic analysis and dual‐luciferase reporter assay were applied to analyse molecular interaction. Next, we conducted immunofluorescence and Western blot for mechanic investigation. Last but not the least, xenograft tumours in nude mice were built by subcutaneously injecting Hep3B cells stably transfected with sh‐NC and sh‐DANCR to detect proliferation and SMMC‐7721 cells stably transfected with sh‐NC and sh‐DANCR to investigate metastasis. RESULTS: The results of qRT‐PCR and bioinformatic analysis revealed the high expression of DANCR in HCC. DANCR accelerated proliferation and metastasis of HCC cells and the knockdown of DANCR had the opposite effect. Meanwhile, xenograft tumours in sh‐DANCR group grow slower and have smaller volumes compared with negative control group. Next, the antineoplastic effect of miR‐27a‐3p on cell growth and motility of HCC was confirmed. In addition, we clarified that DANCR acted as a ceRNA to decoy miR‐27a‐3p via mediating ROCK1/LIMK1/COFILIN1 pathway. In the end, we validated that DANCR/miR‐27a‐3p axis regulates EMT progression by cell immunofluorescence and Western blot. CONCLUSIONS: In a word, DANCR promotes HCC development and induces EMT by decoying miR‐27a‐3p to regulate ROCK1/LIMK1/COFILIN1 pathway. |
format | Online Article Text |
id | pubmed-6668976 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-66689762020-03-13 DANCR promotes HCC progression and regulates EMT by sponging miR‐27a‐3p via ROCK1/LIMK1/COFILIN1 pathway Guo, Dan Li, Yarui Chen, Yifei Zhang, Dan Wang, Xin Lu, Guifang Ren, Mudan Lu, Xinlan He, Shuixiang Cell Prolif Original Articles OBJECTIVES: This research aims to verify that the long non‐coding RNA differentiation antagonizing nonprotein coding RNA (LncRNA DANCR) could modulate the proliferation and metastasis of hepatocellular carcinoma (HCC), and it thus may work as a novel biomarker to render new orientation for early diagnosis and clinical therapy of HCC. MATERIALS AND METHODS: Firstly, qRT‐PCR was used to detect the expression of genes including LncRNA DANCR and miR‐27a‐3p. Next, MTT assay, Ethynyldeoxyuridine (EdU) analysis and clone formation assay were used for investigating cell growth and proliferation. Meanwhile, transwell assay and wound healing assay were applied to evaluate the capacity of cell metastasis and motility, respectively. In addition, bioinformatic analysis and dual‐luciferase reporter assay were applied to analyse molecular interaction. Next, we conducted immunofluorescence and Western blot for mechanic investigation. Last but not the least, xenograft tumours in nude mice were built by subcutaneously injecting Hep3B cells stably transfected with sh‐NC and sh‐DANCR to detect proliferation and SMMC‐7721 cells stably transfected with sh‐NC and sh‐DANCR to investigate metastasis. RESULTS: The results of qRT‐PCR and bioinformatic analysis revealed the high expression of DANCR in HCC. DANCR accelerated proliferation and metastasis of HCC cells and the knockdown of DANCR had the opposite effect. Meanwhile, xenograft tumours in sh‐DANCR group grow slower and have smaller volumes compared with negative control group. Next, the antineoplastic effect of miR‐27a‐3p on cell growth and motility of HCC was confirmed. In addition, we clarified that DANCR acted as a ceRNA to decoy miR‐27a‐3p via mediating ROCK1/LIMK1/COFILIN1 pathway. In the end, we validated that DANCR/miR‐27a‐3p axis regulates EMT progression by cell immunofluorescence and Western blot. CONCLUSIONS: In a word, DANCR promotes HCC development and induces EMT by decoying miR‐27a‐3p to regulate ROCK1/LIMK1/COFILIN1 pathway. John Wiley and Sons Inc. 2019-04-30 /pmc/articles/PMC6668976/ /pubmed/31038266 http://dx.doi.org/10.1111/cpr.12628 Text en © 2019 The Authors. Cell Proliferation Published by John Wiley & Sons Ltd This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Articles Guo, Dan Li, Yarui Chen, Yifei Zhang, Dan Wang, Xin Lu, Guifang Ren, Mudan Lu, Xinlan He, Shuixiang DANCR promotes HCC progression and regulates EMT by sponging miR‐27a‐3p via ROCK1/LIMK1/COFILIN1 pathway |
title | DANCR promotes HCC progression and regulates EMT by sponging miR‐27a‐3p via ROCK1/LIMK1/COFILIN1 pathway |
title_full | DANCR promotes HCC progression and regulates EMT by sponging miR‐27a‐3p via ROCK1/LIMK1/COFILIN1 pathway |
title_fullStr | DANCR promotes HCC progression and regulates EMT by sponging miR‐27a‐3p via ROCK1/LIMK1/COFILIN1 pathway |
title_full_unstemmed | DANCR promotes HCC progression and regulates EMT by sponging miR‐27a‐3p via ROCK1/LIMK1/COFILIN1 pathway |
title_short | DANCR promotes HCC progression and regulates EMT by sponging miR‐27a‐3p via ROCK1/LIMK1/COFILIN1 pathway |
title_sort | dancr promotes hcc progression and regulates emt by sponging mir‐27a‐3p via rock1/limk1/cofilin1 pathway |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6668976/ https://www.ncbi.nlm.nih.gov/pubmed/31038266 http://dx.doi.org/10.1111/cpr.12628 |
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