Project description:Tumour hypoxia is a recognised driver of breast cancer pathology. The main cellular response to hypoxia is mediated by the hypoxia-inducible factors HIF1 and HIF2, and is controlled through the regulation of oxygen-labile HIFα subunits. HIF1α has a well-established role in breast cancer where it has also been shown to be regulated by growth factor signalling. However, the role of HIF2α has been less thoroughly researched. Here, the role of HIF2α was investigated in breast cancer cell lines and publicly available gene expression datasets to determine its relationship with HER2 receptor expression in breast cancer. Using an isogenic cell line model for HER2 overexpression, we establish a direct role for HER2 in driving HIF2α expression in breast cancer. The effect of HER2-mediated HIF2α expression on the cellular response to acute and chronic hypoxia was investigated in 2D and 3D cell line models, and through protein and gene expression analysis, HER2 was shown to drive an exacerbated hypoxic response in these cells. In growth assays, HER2-overexpressing cell lines were shown to be highly sensitive to HIF2-specific inhibition through HIF2α-targeted siRNA and treatment with the HIF2α-specific translation inhibitor C76. Additionally, survival analysis in a large, publicly available dataset demonstrated a relationship between HIF2α and poor disease-specific survival in HER2-overexpressing tumours. We demonstrate a novel role for HIF2α in driving an increased hypoxic response in breast cancer cells and suggest HIF2 signalling may be an important, targetable pathway in HER2-positive breast cancers.
Project description:miRNA expression is often disregulated in cancer pathways, such as progression of resistance. We used microarrays in order to determine the differences in miRNA expression between breast cancer cell lines and their resistant pairs.
Project description:Patient-derived primary HER2+ Leptomeningeal carcinomatosis (Lepto) cell lines were established. RNA-seq in various breast cancer cell lines were conducted to characterize the established cell lines. Gene expression analysis clearly indicated thatt Lepto cell lines are transcriptionally different from HER2 positive and negative metastatic breast cancer cell lines such as BT474 and MDA-MB-231 cells.
Project description:Thirteen HER2 positive breast cancer cell lines were screened with 22 commercially available compounds, mainly targeting proteins in the ErbB2 signaling pathway, and the molecular mechanisms related to treatment response were sought. To search for response predictors, genomic and transcriptomic profiling, PIK3CA mutations and PTEN status were associated to the drug responses and several genes involved in the response of the compounds were identified. Array-CGH experiments of HER2+ breast cancer cell lines grown under standard conditions. DNA from four HER2 positive breast cancer cell lines was isolated and hybridized on Agilent arrays.
Project description:Thirteen HER2 positive breast cancer cell lines were screened with 22 commercially available compounds, mainly targeting proteins in the ErbB2 signaling pathway, and the molecular mechanisms related to treatment response were sought. To search for response predictors, genomic and transcriptomic profiling, PIK3CA mutations and PTEN status were associated to the drug responses and several genes involved in the response of the compounds were identified. RNA from thirteen HER2 positive breast cancer cell lines was isolated and hybridized on Affymetrix arrays.
Project description:Array-CGH experiments of HER2+ breast cancer cell lines grown under standard conditions. All hybridizations performed using pooled normal female DNA as reference. 11 cell lines profiled in total, 10 HER2+, 1 HER2-. Each was hybridized once on an Agilent 244k aCGH microarray.
Project description:Thirteen HER2 positive breast cancer cell lines were screened with 22 commercially available compounds, mainly targeting proteins in the ErbB2 signaling pathway, and the molecular mechanisms related to treatment response were sought. To search for response predictors, genomic and transcriptomic profiling, PIK3CA mutations and PTEN status were associated to the drug responses and several genes involved in the response of the compounds were identified. Array-CGH experiments of HER2+ breast cancer cell lines grown under standard conditions.
Project description:Our findings demonstrate that CDCP1 is a novel modulator of HER2 signalling, and a biomarker for the stratification of breast cancer patients with poor prognosis GEP analysis of human breast cancer cell lines SKBR3 overexpressing CDCP1 and control.
Project description:In the normal breast, PMCA2 transports calcium into milk. It is also upregulated in breast cancer cells and high tumor levels predict increased mortality in patients. In this study, we examined interactions between PMCA2 and HER2. We find that PMCA2 and a scaffolding protein, NHERF1, interact together with HER2 in specific membrane domains protruding from the surface of breast cancer cells. Knocking down PMCA2 or NHERF1 increases intracellular calcium and leads to the ubiquitination, endocytosis and degradation of HER2 after receptor activation. This is associated with reductions in HER2 expression and signaling. Manipulating PMCA2 levels regulates proliferation and apoptosis of breast cancer cells in vitro and knocking out PMCA2 dramatically inhibits the formation of hyperplasia and tumors in MMTV-Neu mice. Gene expression profiling was performed on control SKBR3 cells vs. PMCA2, NHERFR1 and HER2 knock down cells. Each group has 6 duplicates.