Cargando…
Downregulation of miRNA-214 in cancer-associated fibroblasts contributes to migration and invasion of gastric cancer cells through targeting FGF9 and inducing EMT
BACKGROUND: Cancer-associated fibroblasts (CAFs), one of the principal constituents of the tumor microenvironment, have a pivotal role in tumor progression. Dysregulation of microRNAs (miRNAs) in CAFs contributes to the tumor-promoting ability of CAFs. However, the mechanism underlying the involveme...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6334467/ https://www.ncbi.nlm.nih.gov/pubmed/30646925 http://dx.doi.org/10.1186/s13046-018-0995-9 |
_version_ | 1783387723328913408 |
---|---|
author | Wang, Ruifen Sun, Yeqi Yu, Wenwei Yan, Yu Qiao, Meng Jiang, Ruiqi Guan, Wenbin Wang, Lifeng |
author_facet | Wang, Ruifen Sun, Yeqi Yu, Wenwei Yan, Yu Qiao, Meng Jiang, Ruiqi Guan, Wenbin Wang, Lifeng |
author_sort | Wang, Ruifen |
collection | PubMed |
description | BACKGROUND: Cancer-associated fibroblasts (CAFs), one of the principal constituents of the tumor microenvironment, have a pivotal role in tumor progression. Dysregulation of microRNAs (miRNAs) in CAFs contributes to the tumor-promoting ability of CAFs. However, the mechanism underlying the involvement of miRNAs in CAFs of gastric cancer (GC) is not fully understood. This study aimed to explore the effects of miRNA-214 in CAFs on GC migration and invasion. METHODS: The primary CAFs and corresponding normal fibroblasts (NFs) were isolated. Cell counting kit-8, EdU cell proliferation staining and Transwell assays were used to determine the role of miRNA-214 in GC progression. Real-time polymerase chain reaction, Western blot analysis, and dual-luciferase reporter assay were performed to verify the target genes of miRNA-214. Immunofluorescence and Western blot analysis were applied to detect the expression of epithelial–mesenchymal transition (EMT) markers. Immunohistochemistry and in situ hybridization were implemented to analyze the fibroblast growth factor 9 (FGF9) and miRNA-214 expression in human GC tissues, respectively. Finally, to assess its prognostic relevance, Kaplan–Meier survival analysis was conducted. RESULTS: MiRNA-214 was significantly downregulated in CAFs of GC compared with NFs. The upregulation of miRNA-214 in CAFs inhibited GC cell migration and invasion in vitro but failed to affect proliferation. Moreover, GC cells cultured with conditioned medium from CAFs transfected with miR-214 mimic showed increased expression of E-cadherin and decreased expression of Vimentin, N-cadherin and Snail, indicating the suppression of EMT of GC cells. Furthermore, FGF9 was proved to be a direct target gene of miR-214. The expression of FGF9 was higher in CAFs than that in tumor cells not only in primary tumor but also in lymph node metastatic sites (30.0% vs 11.9%, P < 0.01 and 32.1% vs 12.3%, P < 0.01, respectively). Abnormal expression of FGF9 in CAFs of lymph node metastatic sites was significantly associated with poor prognosis in patients with GC (P < 0.05). CONCLUSIONS: This study showed that miR-214 inhibited the tumor-promoting effect of CAFs on GC through targeting FGF9 in CAFs and regulating the EMT process in GC cells, suggesting miRNA-214/FGF9 in CAFs as a potential target for therapeutic approaches in GC. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13046-018-0995-9) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6334467 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-63344672019-01-23 Downregulation of miRNA-214 in cancer-associated fibroblasts contributes to migration and invasion of gastric cancer cells through targeting FGF9 and inducing EMT Wang, Ruifen Sun, Yeqi Yu, Wenwei Yan, Yu Qiao, Meng Jiang, Ruiqi Guan, Wenbin Wang, Lifeng J Exp Clin Cancer Res Research BACKGROUND: Cancer-associated fibroblasts (CAFs), one of the principal constituents of the tumor microenvironment, have a pivotal role in tumor progression. Dysregulation of microRNAs (miRNAs) in CAFs contributes to the tumor-promoting ability of CAFs. However, the mechanism underlying the involvement of miRNAs in CAFs of gastric cancer (GC) is not fully understood. This study aimed to explore the effects of miRNA-214 in CAFs on GC migration and invasion. METHODS: The primary CAFs and corresponding normal fibroblasts (NFs) were isolated. Cell counting kit-8, EdU cell proliferation staining and Transwell assays were used to determine the role of miRNA-214 in GC progression. Real-time polymerase chain reaction, Western blot analysis, and dual-luciferase reporter assay were performed to verify the target genes of miRNA-214. Immunofluorescence and Western blot analysis were applied to detect the expression of epithelial–mesenchymal transition (EMT) markers. Immunohistochemistry and in situ hybridization were implemented to analyze the fibroblast growth factor 9 (FGF9) and miRNA-214 expression in human GC tissues, respectively. Finally, to assess its prognostic relevance, Kaplan–Meier survival analysis was conducted. RESULTS: MiRNA-214 was significantly downregulated in CAFs of GC compared with NFs. The upregulation of miRNA-214 in CAFs inhibited GC cell migration and invasion in vitro but failed to affect proliferation. Moreover, GC cells cultured with conditioned medium from CAFs transfected with miR-214 mimic showed increased expression of E-cadherin and decreased expression of Vimentin, N-cadherin and Snail, indicating the suppression of EMT of GC cells. Furthermore, FGF9 was proved to be a direct target gene of miR-214. The expression of FGF9 was higher in CAFs than that in tumor cells not only in primary tumor but also in lymph node metastatic sites (30.0% vs 11.9%, P < 0.01 and 32.1% vs 12.3%, P < 0.01, respectively). Abnormal expression of FGF9 in CAFs of lymph node metastatic sites was significantly associated with poor prognosis in patients with GC (P < 0.05). CONCLUSIONS: This study showed that miR-214 inhibited the tumor-promoting effect of CAFs on GC through targeting FGF9 in CAFs and regulating the EMT process in GC cells, suggesting miRNA-214/FGF9 in CAFs as a potential target for therapeutic approaches in GC. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13046-018-0995-9) contains supplementary material, which is available to authorized users. BioMed Central 2019-01-15 /pmc/articles/PMC6334467/ /pubmed/30646925 http://dx.doi.org/10.1186/s13046-018-0995-9 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, Ruifen Sun, Yeqi Yu, Wenwei Yan, Yu Qiao, Meng Jiang, Ruiqi Guan, Wenbin Wang, Lifeng Downregulation of miRNA-214 in cancer-associated fibroblasts contributes to migration and invasion of gastric cancer cells through targeting FGF9 and inducing EMT |
title | Downregulation of miRNA-214 in cancer-associated fibroblasts contributes to migration and invasion of gastric cancer cells through targeting FGF9 and inducing EMT |
title_full | Downregulation of miRNA-214 in cancer-associated fibroblasts contributes to migration and invasion of gastric cancer cells through targeting FGF9 and inducing EMT |
title_fullStr | Downregulation of miRNA-214 in cancer-associated fibroblasts contributes to migration and invasion of gastric cancer cells through targeting FGF9 and inducing EMT |
title_full_unstemmed | Downregulation of miRNA-214 in cancer-associated fibroblasts contributes to migration and invasion of gastric cancer cells through targeting FGF9 and inducing EMT |
title_short | Downregulation of miRNA-214 in cancer-associated fibroblasts contributes to migration and invasion of gastric cancer cells through targeting FGF9 and inducing EMT |
title_sort | downregulation of mirna-214 in cancer-associated fibroblasts contributes to migration and invasion of gastric cancer cells through targeting fgf9 and inducing emt |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6334467/ https://www.ncbi.nlm.nih.gov/pubmed/30646925 http://dx.doi.org/10.1186/s13046-018-0995-9 |
work_keys_str_mv | AT wangruifen downregulationofmirna214incancerassociatedfibroblastscontributestomigrationandinvasionofgastriccancercellsthroughtargetingfgf9andinducingemt AT sunyeqi downregulationofmirna214incancerassociatedfibroblastscontributestomigrationandinvasionofgastriccancercellsthroughtargetingfgf9andinducingemt AT yuwenwei downregulationofmirna214incancerassociatedfibroblastscontributestomigrationandinvasionofgastriccancercellsthroughtargetingfgf9andinducingemt AT yanyu downregulationofmirna214incancerassociatedfibroblastscontributestomigrationandinvasionofgastriccancercellsthroughtargetingfgf9andinducingemt AT qiaomeng downregulationofmirna214incancerassociatedfibroblastscontributestomigrationandinvasionofgastriccancercellsthroughtargetingfgf9andinducingemt AT jiangruiqi downregulationofmirna214incancerassociatedfibroblastscontributestomigrationandinvasionofgastriccancercellsthroughtargetingfgf9andinducingemt AT guanwenbin downregulationofmirna214incancerassociatedfibroblastscontributestomigrationandinvasionofgastriccancercellsthroughtargetingfgf9andinducingemt AT wanglifeng downregulationofmirna214incancerassociatedfibroblastscontributestomigrationandinvasionofgastriccancercellsthroughtargetingfgf9andinducingemt |