Cargando…
Tumor-associated macrophage-derived IL-6 and IL-8 enhance invasive activity of LoVo cells induced by PRL-3 in a KCNN4 channel-dependent manner
BACKGROUND: Tumor-associated macrophages (TAMs) are known to promote cancer progression and metastasis through the release of a variety of cytokines. Phosphatase of regenerating liver (PRL-3) has been considered as a marker of colorectal cancer (CRC) liver metastasis. Our previous research suggests...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2014
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4024187/ https://www.ncbi.nlm.nih.gov/pubmed/24885636 http://dx.doi.org/10.1186/1471-2407-14-330 |
_version_ | 1782316630353641472 |
---|---|
author | Xu, Heyang Lai, Wei Zhang, Yang Liu, Lu Luo, Xingxi Zeng, Yujie Wu, Heng Lan, Qiusheng Chu, Zhonghua |
author_facet | Xu, Heyang Lai, Wei Zhang, Yang Liu, Lu Luo, Xingxi Zeng, Yujie Wu, Heng Lan, Qiusheng Chu, Zhonghua |
author_sort | Xu, Heyang |
collection | PubMed |
description | BACKGROUND: Tumor-associated macrophages (TAMs) are known to promote cancer progression and metastasis through the release of a variety of cytokines. Phosphatase of regenerating liver (PRL-3) has been considered as a marker of colorectal cancer (CRC) liver metastasis. Our previous research suggests that PRL-3 can enhance the metastasis of CRC through the up-regulation of intermediate-conductance Ca(2+)-activated K(+) (KCNN4) channel, which is dependent on the autocrine secretion of tumor necrosis factor-alpha (TNF-α). However, whether TAMs participate in the progression and metastasis of CRC induced by PRL-3 remains unknown. METHODS: We used flow cytometry, coculture, western blotting, invasion assays, real-time quantitative PCR, chromatin immunoprecipitation, luciferase reporter assays, and immunofluorescence staining to determine the effect of TAMs on the ability of PRL-3 to promote invasiveness of CRC cells. RESULTS: In this study, we found that TAMs facilitated the metastasis of CRC induced by PRL-3. When TAMs were cocultured with CRC cells, the expression of KCNN4 was increased in TAMs and the invasion of CRC cells was enhanced. Furthermore, cytokines that were secreted by TAMs, such as IL-6 and IL-8, were also significantly increased. This response was attenuated by treating TAMs with the KCNN4 channel-specific inhibitor, 1-[(2-chlorophenyl) diphenylmethyl]-1H-pyrazole (TRAM-34), which suggested that KCNN4 channels may be involved in inducing the secretion of IL-6 and IL-8 by TAMs and improving CRC cell invasiveness. Moreover, the expression of KCNN4 channels in TAMs was regulated through the NF-κB signal pathway, which is activated by TNF-α from CRC cells. Immunofluorescence analysis of colorectal specimens indicated that IL-6 and IL-8 double positive cells in the stroma showed positive staining for the TAM marker CD68, suggesting that TAMs produce IL-6 and IL-8. Increased numbers of these cells correlated with higher clinical stage. CONCLUSIONS: Our findings suggested that TAMs participate in the metastasis of CRC induced by PRL-3 through the TNF-α mediated secretion of IL-6 and IL-8 in a paracrine manner. |
format | Online Article Text |
id | pubmed-4024187 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-40241872014-05-18 Tumor-associated macrophage-derived IL-6 and IL-8 enhance invasive activity of LoVo cells induced by PRL-3 in a KCNN4 channel-dependent manner Xu, Heyang Lai, Wei Zhang, Yang Liu, Lu Luo, Xingxi Zeng, Yujie Wu, Heng Lan, Qiusheng Chu, Zhonghua BMC Cancer Research Article BACKGROUND: Tumor-associated macrophages (TAMs) are known to promote cancer progression and metastasis through the release of a variety of cytokines. Phosphatase of regenerating liver (PRL-3) has been considered as a marker of colorectal cancer (CRC) liver metastasis. Our previous research suggests that PRL-3 can enhance the metastasis of CRC through the up-regulation of intermediate-conductance Ca(2+)-activated K(+) (KCNN4) channel, which is dependent on the autocrine secretion of tumor necrosis factor-alpha (TNF-α). However, whether TAMs participate in the progression and metastasis of CRC induced by PRL-3 remains unknown. METHODS: We used flow cytometry, coculture, western blotting, invasion assays, real-time quantitative PCR, chromatin immunoprecipitation, luciferase reporter assays, and immunofluorescence staining to determine the effect of TAMs on the ability of PRL-3 to promote invasiveness of CRC cells. RESULTS: In this study, we found that TAMs facilitated the metastasis of CRC induced by PRL-3. When TAMs were cocultured with CRC cells, the expression of KCNN4 was increased in TAMs and the invasion of CRC cells was enhanced. Furthermore, cytokines that were secreted by TAMs, such as IL-6 and IL-8, were also significantly increased. This response was attenuated by treating TAMs with the KCNN4 channel-specific inhibitor, 1-[(2-chlorophenyl) diphenylmethyl]-1H-pyrazole (TRAM-34), which suggested that KCNN4 channels may be involved in inducing the secretion of IL-6 and IL-8 by TAMs and improving CRC cell invasiveness. Moreover, the expression of KCNN4 channels in TAMs was regulated through the NF-κB signal pathway, which is activated by TNF-α from CRC cells. Immunofluorescence analysis of colorectal specimens indicated that IL-6 and IL-8 double positive cells in the stroma showed positive staining for the TAM marker CD68, suggesting that TAMs produce IL-6 and IL-8. Increased numbers of these cells correlated with higher clinical stage. CONCLUSIONS: Our findings suggested that TAMs participate in the metastasis of CRC induced by PRL-3 through the TNF-α mediated secretion of IL-6 and IL-8 in a paracrine manner. BioMed Central 2014-05-10 /pmc/articles/PMC4024187/ /pubmed/24885636 http://dx.doi.org/10.1186/1471-2407-14-330 Text en Copyright © 2014 Xu et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. 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 Article Xu, Heyang Lai, Wei Zhang, Yang Liu, Lu Luo, Xingxi Zeng, Yujie Wu, Heng Lan, Qiusheng Chu, Zhonghua Tumor-associated macrophage-derived IL-6 and IL-8 enhance invasive activity of LoVo cells induced by PRL-3 in a KCNN4 channel-dependent manner |
title | Tumor-associated macrophage-derived IL-6 and IL-8 enhance invasive activity of LoVo cells induced by PRL-3 in a KCNN4 channel-dependent manner |
title_full | Tumor-associated macrophage-derived IL-6 and IL-8 enhance invasive activity of LoVo cells induced by PRL-3 in a KCNN4 channel-dependent manner |
title_fullStr | Tumor-associated macrophage-derived IL-6 and IL-8 enhance invasive activity of LoVo cells induced by PRL-3 in a KCNN4 channel-dependent manner |
title_full_unstemmed | Tumor-associated macrophage-derived IL-6 and IL-8 enhance invasive activity of LoVo cells induced by PRL-3 in a KCNN4 channel-dependent manner |
title_short | Tumor-associated macrophage-derived IL-6 and IL-8 enhance invasive activity of LoVo cells induced by PRL-3 in a KCNN4 channel-dependent manner |
title_sort | tumor-associated macrophage-derived il-6 and il-8 enhance invasive activity of lovo cells induced by prl-3 in a kcnn4 channel-dependent manner |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4024187/ https://www.ncbi.nlm.nih.gov/pubmed/24885636 http://dx.doi.org/10.1186/1471-2407-14-330 |
work_keys_str_mv | AT xuheyang tumorassociatedmacrophagederivedil6andil8enhanceinvasiveactivityoflovocellsinducedbyprl3inakcnn4channeldependentmanner AT laiwei tumorassociatedmacrophagederivedil6andil8enhanceinvasiveactivityoflovocellsinducedbyprl3inakcnn4channeldependentmanner AT zhangyang tumorassociatedmacrophagederivedil6andil8enhanceinvasiveactivityoflovocellsinducedbyprl3inakcnn4channeldependentmanner AT liulu tumorassociatedmacrophagederivedil6andil8enhanceinvasiveactivityoflovocellsinducedbyprl3inakcnn4channeldependentmanner AT luoxingxi tumorassociatedmacrophagederivedil6andil8enhanceinvasiveactivityoflovocellsinducedbyprl3inakcnn4channeldependentmanner AT zengyujie tumorassociatedmacrophagederivedil6andil8enhanceinvasiveactivityoflovocellsinducedbyprl3inakcnn4channeldependentmanner AT wuheng tumorassociatedmacrophagederivedil6andil8enhanceinvasiveactivityoflovocellsinducedbyprl3inakcnn4channeldependentmanner AT lanqiusheng tumorassociatedmacrophagederivedil6andil8enhanceinvasiveactivityoflovocellsinducedbyprl3inakcnn4channeldependentmanner AT chuzhonghua tumorassociatedmacrophagederivedil6andil8enhanceinvasiveactivityoflovocellsinducedbyprl3inakcnn4channeldependentmanner |