Cargando…
MED12 exerts an emerging role in actin-mediated cytokinesis via LIMK2/cofilin pathway in NSCLC
BACKGROUND: Mediator complex subunit 12 (MED12) is an essential hub for transcriptional regulation, in which mutations and overexpression were reported to be associated with several kinds of malignancies. Nevertheless, the role of MED12 in non-small cell lung cancer (NSCLC) remains to be elucidated....
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6509838/ https://www.ncbi.nlm.nih.gov/pubmed/31072327 http://dx.doi.org/10.1186/s12943-019-1020-4 |
_version_ | 1783417328704159744 |
---|---|
author | Xu, Meng Wang, Fang Li, Guibo Wang, Xiaokun Fang, Xiaona Jin, Haoxuan Chen, Zhen Zhang, Jianye Fu, Liwu |
author_facet | Xu, Meng Wang, Fang Li, Guibo Wang, Xiaokun Fang, Xiaona Jin, Haoxuan Chen, Zhen Zhang, Jianye Fu, Liwu |
author_sort | Xu, Meng |
collection | PubMed |
description | BACKGROUND: Mediator complex subunit 12 (MED12) is an essential hub for transcriptional regulation, in which mutations and overexpression were reported to be associated with several kinds of malignancies. Nevertheless, the role of MED12 in non-small cell lung cancer (NSCLC) remains to be elucidated. METHODS: MED12 mutation was detected by Next-generation sequencing. The expression of MED12 in 179 human NSCLC tissue samples and 73 corresponding adjacent normal lung tissue samples was measured by immunohistochemistry (IHC). CRISPR-Cas9 was used to knock out MED12 in PC9 and SPC-A1 cells. MED12 rescued stable cell lines were generated by lentivirus infection. We traced cell division process by live cell imaging. The molecular mechanism of aborted cytokinesis resulted by MED12 knockout was investigated by RNA-seq. Effects of MED12 deletion on the proliferation of NSCLC cells were determined by MTT assay and Colony-formation assay in vitro and xenograft tumor model in nude mouse. Cell senescence was measured by SA-β-gal staining. RESULTS: In our study, no MED12 exon mutation was detected in NSCLC samples, whereas we found that MED12 was overexpressed in human NSCLC tissues, which positively correlated with the tumor volume and adversely affected patient survival. Furthermore, knockout MED12 in NSCLC cell lines resulted in cytokinesis failure, displayed a multinuclear phenotype, and disposed to senescence, and become non-viable. Lack of MED12 decreased the proliferative potential of NSCLC cells and limited the tumor growth in vivo. Mechanism investigations revealed that MED12 knockout activated LIMK2, caused aberrant actin cytoskeleton remodeling, and disrupted the abscission of intercellular bridge, which led to the cytokinesis failure. Reconstitution of exogenous MED12 restored actin dynamics, normal cytokinesis and cell proliferation capacity in MED12 knockout cells. CONCLUSIONS: These results revealed a novel role of MED12 as an important regulator for maintaining accurate cytokinesis and survival in NSCLC cells, which may offer a therapeutic strategy to control tumor growth for NSCLC patients especially those highly expressed MED12. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12943-019-1020-4) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6509838 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-65098382019-06-05 MED12 exerts an emerging role in actin-mediated cytokinesis via LIMK2/cofilin pathway in NSCLC Xu, Meng Wang, Fang Li, Guibo Wang, Xiaokun Fang, Xiaona Jin, Haoxuan Chen, Zhen Zhang, Jianye Fu, Liwu Mol Cancer Research BACKGROUND: Mediator complex subunit 12 (MED12) is an essential hub for transcriptional regulation, in which mutations and overexpression were reported to be associated with several kinds of malignancies. Nevertheless, the role of MED12 in non-small cell lung cancer (NSCLC) remains to be elucidated. METHODS: MED12 mutation was detected by Next-generation sequencing. The expression of MED12 in 179 human NSCLC tissue samples and 73 corresponding adjacent normal lung tissue samples was measured by immunohistochemistry (IHC). CRISPR-Cas9 was used to knock out MED12 in PC9 and SPC-A1 cells. MED12 rescued stable cell lines were generated by lentivirus infection. We traced cell division process by live cell imaging. The molecular mechanism of aborted cytokinesis resulted by MED12 knockout was investigated by RNA-seq. Effects of MED12 deletion on the proliferation of NSCLC cells were determined by MTT assay and Colony-formation assay in vitro and xenograft tumor model in nude mouse. Cell senescence was measured by SA-β-gal staining. RESULTS: In our study, no MED12 exon mutation was detected in NSCLC samples, whereas we found that MED12 was overexpressed in human NSCLC tissues, which positively correlated with the tumor volume and adversely affected patient survival. Furthermore, knockout MED12 in NSCLC cell lines resulted in cytokinesis failure, displayed a multinuclear phenotype, and disposed to senescence, and become non-viable. Lack of MED12 decreased the proliferative potential of NSCLC cells and limited the tumor growth in vivo. Mechanism investigations revealed that MED12 knockout activated LIMK2, caused aberrant actin cytoskeleton remodeling, and disrupted the abscission of intercellular bridge, which led to the cytokinesis failure. Reconstitution of exogenous MED12 restored actin dynamics, normal cytokinesis and cell proliferation capacity in MED12 knockout cells. CONCLUSIONS: These results revealed a novel role of MED12 as an important regulator for maintaining accurate cytokinesis and survival in NSCLC cells, which may offer a therapeutic strategy to control tumor growth for NSCLC patients especially those highly expressed MED12. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12943-019-1020-4) contains supplementary material, which is available to authorized users. BioMed Central 2019-05-09 /pmc/articles/PMC6509838/ /pubmed/31072327 http://dx.doi.org/10.1186/s12943-019-1020-4 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 Xu, Meng Wang, Fang Li, Guibo Wang, Xiaokun Fang, Xiaona Jin, Haoxuan Chen, Zhen Zhang, Jianye Fu, Liwu MED12 exerts an emerging role in actin-mediated cytokinesis via LIMK2/cofilin pathway in NSCLC |
title | MED12 exerts an emerging role in actin-mediated cytokinesis via LIMK2/cofilin pathway in NSCLC |
title_full | MED12 exerts an emerging role in actin-mediated cytokinesis via LIMK2/cofilin pathway in NSCLC |
title_fullStr | MED12 exerts an emerging role in actin-mediated cytokinesis via LIMK2/cofilin pathway in NSCLC |
title_full_unstemmed | MED12 exerts an emerging role in actin-mediated cytokinesis via LIMK2/cofilin pathway in NSCLC |
title_short | MED12 exerts an emerging role in actin-mediated cytokinesis via LIMK2/cofilin pathway in NSCLC |
title_sort | med12 exerts an emerging role in actin-mediated cytokinesis via limk2/cofilin pathway in nsclc |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6509838/ https://www.ncbi.nlm.nih.gov/pubmed/31072327 http://dx.doi.org/10.1186/s12943-019-1020-4 |
work_keys_str_mv | AT xumeng med12exertsanemergingroleinactinmediatedcytokinesisvialimk2cofilinpathwayinnsclc AT wangfang med12exertsanemergingroleinactinmediatedcytokinesisvialimk2cofilinpathwayinnsclc AT liguibo med12exertsanemergingroleinactinmediatedcytokinesisvialimk2cofilinpathwayinnsclc AT wangxiaokun med12exertsanemergingroleinactinmediatedcytokinesisvialimk2cofilinpathwayinnsclc AT fangxiaona med12exertsanemergingroleinactinmediatedcytokinesisvialimk2cofilinpathwayinnsclc AT jinhaoxuan med12exertsanemergingroleinactinmediatedcytokinesisvialimk2cofilinpathwayinnsclc AT chenzhen med12exertsanemergingroleinactinmediatedcytokinesisvialimk2cofilinpathwayinnsclc AT zhangjianye med12exertsanemergingroleinactinmediatedcytokinesisvialimk2cofilinpathwayinnsclc AT fuliwu med12exertsanemergingroleinactinmediatedcytokinesisvialimk2cofilinpathwayinnsclc |