Transcriptomic and ChIP-sequence interrogation of EGFR signaling in HER2+ breast cancer cells reveals a dynamic chromatin landscape and S100 genes as targets
Abstract Background The Human Epidermal Growth Factor Receptor (EGFR/HER1) can be activated by several ligands including Transforming Growth Factor alpha (TGF-α) and Epidermal Growth Factor (EGF). Following ligand binding, EGFR heterodimerizes with other HER family members, such as HER2 (human epide...
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| Format: | Article |
| Language: | English |
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BMC
2019-02-01
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| Series: | BMC Medical Genomics |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12920-019-0477-8 |
| _version_ | 1818967808512360448 |
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| author | Miguel Nava Pranabananda Dutta Nathan R. Zemke Robin Farias-Eisner Jaydutt V. Vadgama Yanyuan Wu |
| author_facet | Miguel Nava Pranabananda Dutta Nathan R. Zemke Robin Farias-Eisner Jaydutt V. Vadgama Yanyuan Wu |
| author_sort | Miguel Nava |
| collection | DOAJ |
| description | Abstract Background The Human Epidermal Growth Factor Receptor (EGFR/HER1) can be activated by several ligands including Transforming Growth Factor alpha (TGF-α) and Epidermal Growth Factor (EGF). Following ligand binding, EGFR heterodimerizes with other HER family members, such as HER2 (human epidermal growth factor receptor-2). Previously, we showed that the EGFR is upregulated in trastuzumab resistant HER2 positive (HER2+) breast cancer cells. This study is aimed to determine the downstream effects on transcription following EGFR upregulation in HER2+ breast cancer cells. Methods RNA-sequence and ChIP-sequence for H3K18ac and H3K27ac (Histone H3 lysine K18 and K27 acetylation) were conducted following an Epidermal Growth Factor (EGF) treatment time course in HER2+ breast cancer cells, SKBR3. The levels of several proteins of interest were confirmed by western blot analysis. The cellular localization of proteins of interest was examined using biochemically fractionated lysates followed by western blot analysis. Results Over the course of 24 h, EGFR stimulation resulted in the modulation of over 4000 transcripts. Moreover, our data demonstrates that EGFR/HER2 signaling regulates the epigenome, with global H3K18ac and H3K27ac oscillating as a function of time following EGF treatment. RNA-sequence data demonstrates the activation of immediate early genes (IEGs) and delayed early genes (DEGs) within 1 h of EGF treatment. More importantly, we have identified members of the S100 (S100 Calcium Binding Protein) gene family as likely direct targets of EGFR signaling as H3K18ac, H3K27ac and pol2 (RNA polymerase II) increase near the transcription start sites of some of these genes. Conclusions Our data suggests that S100 proteins, which act as Ca2+ sensors, could play a role in EGF induced tumor cell growth and metastasis, contribute to trastuzumab resistance and cell migration and that they are likely drug targets in HER2+ breast cancer. |
| first_indexed | 2024-12-20T13:54:41Z |
| format | Article |
| id | doaj.art-7ca0f430d4454feeabb9e98f27d2c9e7 |
| institution | Directory Open Access Journal |
| issn | 1755-8794 |
| language | English |
| last_indexed | 2024-12-20T13:54:41Z |
| publishDate | 2019-02-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Medical Genomics |
| spelling | doaj.art-7ca0f430d4454feeabb9e98f27d2c9e72022-12-21T19:38:28ZengBMCBMC Medical Genomics1755-87942019-02-0112111510.1186/s12920-019-0477-8Transcriptomic and ChIP-sequence interrogation of EGFR signaling in HER2+ breast cancer cells reveals a dynamic chromatin landscape and S100 genes as targetsMiguel Nava0Pranabananda Dutta1Nathan R. Zemke2Robin Farias-Eisner3Jaydutt V. Vadgama4Yanyuan Wu5Division of Cancer Research and Training, Department of Medicine, Charles R. Drew University of Medicine and ScienceDivision of Cancer Research and Training, Department of Medicine, Charles R. Drew University of Medicine and ScienceMolecular Biology Institute, University of CaliforniaJonsson Comprehensive Cancer Center and David Geffen School of Medicine, University of CaliforniaDivision of Cancer Research and Training, Department of Medicine, Charles R. Drew University of Medicine and ScienceDivision of Cancer Research and Training, Department of Medicine, Charles R. Drew University of Medicine and ScienceAbstract Background The Human Epidermal Growth Factor Receptor (EGFR/HER1) can be activated by several ligands including Transforming Growth Factor alpha (TGF-α) and Epidermal Growth Factor (EGF). Following ligand binding, EGFR heterodimerizes with other HER family members, such as HER2 (human epidermal growth factor receptor-2). Previously, we showed that the EGFR is upregulated in trastuzumab resistant HER2 positive (HER2+) breast cancer cells. This study is aimed to determine the downstream effects on transcription following EGFR upregulation in HER2+ breast cancer cells. Methods RNA-sequence and ChIP-sequence for H3K18ac and H3K27ac (Histone H3 lysine K18 and K27 acetylation) were conducted following an Epidermal Growth Factor (EGF) treatment time course in HER2+ breast cancer cells, SKBR3. The levels of several proteins of interest were confirmed by western blot analysis. The cellular localization of proteins of interest was examined using biochemically fractionated lysates followed by western blot analysis. Results Over the course of 24 h, EGFR stimulation resulted in the modulation of over 4000 transcripts. Moreover, our data demonstrates that EGFR/HER2 signaling regulates the epigenome, with global H3K18ac and H3K27ac oscillating as a function of time following EGF treatment. RNA-sequence data demonstrates the activation of immediate early genes (IEGs) and delayed early genes (DEGs) within 1 h of EGF treatment. More importantly, we have identified members of the S100 (S100 Calcium Binding Protein) gene family as likely direct targets of EGFR signaling as H3K18ac, H3K27ac and pol2 (RNA polymerase II) increase near the transcription start sites of some of these genes. Conclusions Our data suggests that S100 proteins, which act as Ca2+ sensors, could play a role in EGF induced tumor cell growth and metastasis, contribute to trastuzumab resistance and cell migration and that they are likely drug targets in HER2+ breast cancer.http://link.springer.com/article/10.1186/s12920-019-0477-8HER2Breast CancerEGFREpigeneticsChromatinNext generation sequencing |
| spellingShingle | Miguel Nava Pranabananda Dutta Nathan R. Zemke Robin Farias-Eisner Jaydutt V. Vadgama Yanyuan Wu Transcriptomic and ChIP-sequence interrogation of EGFR signaling in HER2+ breast cancer cells reveals a dynamic chromatin landscape and S100 genes as targets BMC Medical Genomics HER2 Breast Cancer EGFR Epigenetics Chromatin Next generation sequencing |
| title | Transcriptomic and ChIP-sequence interrogation of EGFR signaling in HER2+ breast cancer cells reveals a dynamic chromatin landscape and S100 genes as targets |
| title_full | Transcriptomic and ChIP-sequence interrogation of EGFR signaling in HER2+ breast cancer cells reveals a dynamic chromatin landscape and S100 genes as targets |
| title_fullStr | Transcriptomic and ChIP-sequence interrogation of EGFR signaling in HER2+ breast cancer cells reveals a dynamic chromatin landscape and S100 genes as targets |
| title_full_unstemmed | Transcriptomic and ChIP-sequence interrogation of EGFR signaling in HER2+ breast cancer cells reveals a dynamic chromatin landscape and S100 genes as targets |
| title_short | Transcriptomic and ChIP-sequence interrogation of EGFR signaling in HER2+ breast cancer cells reveals a dynamic chromatin landscape and S100 genes as targets |
| title_sort | transcriptomic and chip sequence interrogation of egfr signaling in her2 breast cancer cells reveals a dynamic chromatin landscape and s100 genes as targets |
| topic | HER2 Breast Cancer EGFR Epigenetics Chromatin Next generation sequencing |
| url | http://link.springer.com/article/10.1186/s12920-019-0477-8 |
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