Focal adhesion kinase (FAK) activation by estrogens involves GPER in triple-negative breast cancer cells
Abstract Background Focal adhesion kinase (FAK) is a cytoplasmatic protein tyrosine kinase that associates with both integrins and growth factor receptors toward the adhesion, migration and invasion of cancer cells. The G-protein coupled estrogen receptor (GPER) has been involved in the stimulatory...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-02-01
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| Series: | Journal of Experimental & Clinical Cancer Research |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13046-019-1056-8 |
| _version_ | 1828532727366811648 |
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| author | Damiano Cosimo Rigiracciolo Maria Francesca Santolla Rosamaria Lappano Adele Vivacqua Francesca Cirillo Giulia Raffaella Galli Marianna Talia Lucia Muglia Michele Pellegrino Nijiro Nohata Maria Teresa Di Martino Marcello Maggiolini |
| author_facet | Damiano Cosimo Rigiracciolo Maria Francesca Santolla Rosamaria Lappano Adele Vivacqua Francesca Cirillo Giulia Raffaella Galli Marianna Talia Lucia Muglia Michele Pellegrino Nijiro Nohata Maria Teresa Di Martino Marcello Maggiolini |
| author_sort | Damiano Cosimo Rigiracciolo |
| collection | DOAJ |
| description | Abstract Background Focal adhesion kinase (FAK) is a cytoplasmatic protein tyrosine kinase that associates with both integrins and growth factor receptors toward the adhesion, migration and invasion of cancer cells. The G-protein coupled estrogen receptor (GPER) has been involved in the stimulatory action of estrogens in breast tumor. In this study, we have investigated the engagement of FAK by GPER signaling in triple negative breast cancer (TNBC) cells. Methods Publicly available large-scale database and patient data sets derived from “The Cancer Genome Atlas” (TCGA; www.cbioportal.org) were used to assess FAK expression in TNBC, non-TNBC tumors and normal breast tissues. MDA-MB 231 and SUM159 TNBC cells were used as model system. The levels of phosphorylated FAK, other transduction mediators and target genes were detected by western blotting analysis. Focal adhesion assay was carried out in order to determine the focal adhesion points and the formation of focal adhesions (FAs). Luciferase assays were performed to evaluate the promoters activity of c-FOS, EGR1 and CTGF upon GPER activation. The mRNA expression of the aforementioned genes was measured by real time-PCR. Boyden chamber and wound healing assays were used in order to evaluate cell migration. The statistical analysis was performed by ANOVA. Results We first determined by bioinformatic analysis that the mRNA expression levels of the gene encoding FAK, namely PTK2, is higher in TNBC respect to non-TNBC and normal breast tissues. Next, we found that estrogenic GPER signaling triggers Y397 FAK phosphorylation as well as the increase of focal adhesion points (FAs) in TNBC cells. Besides, we ascertained that GPER and FAK activation are involved in the STAT3 nuclear accumulation and gene expression changes. As biological counterpart, we show that FAK inhibition prevents the migration of TNBC cells upon GPER activation. Conclusions The present data provide novel insights regarding the action of FAK in TNBC. Moreover, on the basis of our findings estrogenic GPER signaling may be considered among the transduction mechanisms engaging FAK toward breast cancer progression. |
| first_indexed | 2024-12-11T22:57:19Z |
| format | Article |
| id | doaj.art-ae60ac6c3e34402aae113891fea6ad26 |
| institution | Directory Open Access Journal |
| issn | 1756-9966 |
| language | English |
| last_indexed | 2024-12-11T22:57:19Z |
| publishDate | 2019-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Experimental & Clinical Cancer Research |
| spelling | doaj.art-ae60ac6c3e34402aae113891fea6ad262022-12-22T00:47:12ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662019-02-0138111610.1186/s13046-019-1056-8Focal adhesion kinase (FAK) activation by estrogens involves GPER in triple-negative breast cancer cellsDamiano Cosimo Rigiracciolo0Maria Francesca Santolla1Rosamaria Lappano2Adele Vivacqua3Francesca Cirillo4Giulia Raffaella Galli5Marianna Talia6Lucia Muglia7Michele Pellegrino8Nijiro Nohata9Maria Teresa Di Martino10Marcello Maggiolini11Department of Pharmacy, Health and Nutritional Sciences, University of CalabriaDepartment of Pharmacy, Health and Nutritional Sciences, University of CalabriaDepartment of Pharmacy, Health and Nutritional Sciences, University of CalabriaDepartment of Pharmacy, Health and Nutritional Sciences, University of CalabriaDepartment of Pharmacy, Health and Nutritional Sciences, University of CalabriaDepartment of Pharmacy, Health and Nutritional Sciences, University of CalabriaDepartment of Pharmacy, Health and Nutritional Sciences, University of CalabriaDepartment of Pharmacy, Health and Nutritional Sciences, University of CalabriaDepartment of Pharmacy, Health and Nutritional Sciences, University of CalabriaMSD K.KDepartment of Experimental and Clinical Medicine, Magna Graecia UniversityDepartment of Pharmacy, Health and Nutritional Sciences, University of CalabriaAbstract Background Focal adhesion kinase (FAK) is a cytoplasmatic protein tyrosine kinase that associates with both integrins and growth factor receptors toward the adhesion, migration and invasion of cancer cells. The G-protein coupled estrogen receptor (GPER) has been involved in the stimulatory action of estrogens in breast tumor. In this study, we have investigated the engagement of FAK by GPER signaling in triple negative breast cancer (TNBC) cells. Methods Publicly available large-scale database and patient data sets derived from “The Cancer Genome Atlas” (TCGA; www.cbioportal.org) were used to assess FAK expression in TNBC, non-TNBC tumors and normal breast tissues. MDA-MB 231 and SUM159 TNBC cells were used as model system. The levels of phosphorylated FAK, other transduction mediators and target genes were detected by western blotting analysis. Focal adhesion assay was carried out in order to determine the focal adhesion points and the formation of focal adhesions (FAs). Luciferase assays were performed to evaluate the promoters activity of c-FOS, EGR1 and CTGF upon GPER activation. The mRNA expression of the aforementioned genes was measured by real time-PCR. Boyden chamber and wound healing assays were used in order to evaluate cell migration. The statistical analysis was performed by ANOVA. Results We first determined by bioinformatic analysis that the mRNA expression levels of the gene encoding FAK, namely PTK2, is higher in TNBC respect to non-TNBC and normal breast tissues. Next, we found that estrogenic GPER signaling triggers Y397 FAK phosphorylation as well as the increase of focal adhesion points (FAs) in TNBC cells. Besides, we ascertained that GPER and FAK activation are involved in the STAT3 nuclear accumulation and gene expression changes. As biological counterpart, we show that FAK inhibition prevents the migration of TNBC cells upon GPER activation. Conclusions The present data provide novel insights regarding the action of FAK in TNBC. Moreover, on the basis of our findings estrogenic GPER signaling may be considered among the transduction mechanisms engaging FAK toward breast cancer progression.http://link.springer.com/article/10.1186/s13046-019-1056-8TNBCMDA-MB 231SUM159GPERG-15FAK |
| spellingShingle | Damiano Cosimo Rigiracciolo Maria Francesca Santolla Rosamaria Lappano Adele Vivacqua Francesca Cirillo Giulia Raffaella Galli Marianna Talia Lucia Muglia Michele Pellegrino Nijiro Nohata Maria Teresa Di Martino Marcello Maggiolini Focal adhesion kinase (FAK) activation by estrogens involves GPER in triple-negative breast cancer cells Journal of Experimental & Clinical Cancer Research TNBC MDA-MB 231 SUM159 GPER G-15 FAK |
| title | Focal adhesion kinase (FAK) activation by estrogens involves GPER in triple-negative breast cancer cells |
| title_full | Focal adhesion kinase (FAK) activation by estrogens involves GPER in triple-negative breast cancer cells |
| title_fullStr | Focal adhesion kinase (FAK) activation by estrogens involves GPER in triple-negative breast cancer cells |
| title_full_unstemmed | Focal adhesion kinase (FAK) activation by estrogens involves GPER in triple-negative breast cancer cells |
| title_short | Focal adhesion kinase (FAK) activation by estrogens involves GPER in triple-negative breast cancer cells |
| title_sort | focal adhesion kinase fak activation by estrogens involves gper in triple negative breast cancer cells |
| topic | TNBC MDA-MB 231 SUM159 GPER G-15 FAK |
| url | http://link.springer.com/article/10.1186/s13046-019-1056-8 |
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