Journal: Cancers

Article Title: Combined Targeting of Estrogen Receptor Alpha and Exportin 1 in Metastatic Breast Cancers

doi: 10.3390/cancers12092397

Figure Lengend Snippet: Clinical relevance of combining XPO1 inhibitors with current therapies in metastatic ER + tumors: Impact on metastasis related gene expression and therapy resistant cells. ( A ) Correlation between XPO1 and PIK3CA expression in ER + tumors in METABRIC and TCGA datasets. Spearman and Pearson correlation analyses coefficients and p -values were indicated for each dataset (METABRIC data set: Spearman correlation = 0.35, Pearson correlation = 0.37, TCGA data set: Spearman correlation = 0.5, Pearson correlation = 0.44). ( B ) RNA-Seq analysis in MCF7- ESR1 Y537S cells, treated with 1 µM 4-OHT, Fulv or Palb in the presence or absence of 100 nM SEL. PCA plot showing distinct gene expression patterns with the different treatments are shown at the right bottom panel of (B). ( C ) Gene-set enrichment analysis (GSEA) of gene sets that were enriched in 4-OHT + SEL dataset. ( D ) Cell viability assay in different endocrine-sensitive or endocrine-resistant breast cancer cell lines (MCF7 Parental, MCF7- ESR1 Y537S , MCF7- ESR1 D538G , T47D Parental, T47D- ESR1 Y537S , T47D- ESR1 D538G , MCF7 cells treated with different treatments and BT474), cotreated with 4-OHT [10 −6 M], Fulv [10 −6 M] and Palb [10 −6 M] and SEL [10 −7 M]. A one-way analysis of variance (ANOVA) model was used for statistical significance of change in cell viability with different treatments. All values were presented as mean ± SEM from six independent repeats. ( E ) Scanning electron microscopy (SEM) images of human tissues and decellularized tissue from which ECM hydrogels are derived. ( F ) Stiffness of TissueSpec ® ECM Hydrogels. ( G ). MCF7- ESR1 Y537S cells were cultured at a density of 2 × 10 3 cells/well and embedded in 3D hydrogels to mimic different metastatic niches (bone, liver, lung) for breast cancer cells, and they were treated with different endocrine agents (4OHT [10 −6 ], Fulv [10 −6 ] and Palb [10 −6 ]) and in combination with SEL [10 −7 ]. ( H ). Colony size (pixels) and number were quantified using OpenCFU colony counting software ( http://opencfu.sourceforge.net/ ). A one-way analysis of variance (ANOVA) model was used for statistical significance of treatment and values were presented as mean ± SEM from three biological replicates (** p <0.01, *** p <0.001; **** p < 0.0001).

Article Snippet: Stiffnesses of TissueSpec ® Bone, Liver, and Lung ECM Hydrogels (#MTSMS10, IN SITE TM Metastasis ECM Hydrogel Kit, Xylyx Bio, Brooklyn, NY, USA) were determined by rheometry analysis.

Techniques: Gene Expression, Expressing, RNA Sequencing, Viability Assay, Electron Microscopy, Derivative Assay, Cell Culture, Software

Journal: Cancers

Article Title: Combined Targeting of Estrogen Receptor Alpha and Exportin 1 in Metastatic Breast Cancers

doi: 10.3390/cancers12092397

Figure Lengend Snippet: Clinical relevance of combining XPO1 inhibitors with current therapies in metastatic ER + tumors: Impact on metastasis related gene expression and therapy resistant cells. ( A ) Correlation between XPO1 and PIK3CA expression in ER + tumors in METABRIC and TCGA datasets. Spearman and Pearson correlation analyses coefficients and p -values were indicated for each dataset (METABRIC data set: Spearman correlation = 0.35, Pearson correlation = 0.37, TCGA data set: Spearman correlation = 0.5, Pearson correlation = 0.44). ( B ) RNA-Seq analysis in MCF7- ESR1 Y537S cells, treated with 1 µM 4-OHT, Fulv or Palb in the presence or absence of 100 nM SEL. PCA plot showing distinct gene expression patterns with the different treatments are shown at the right bottom panel of (B). ( C ) Gene-set enrichment analysis (GSEA) of gene sets that were enriched in 4-OHT + SEL dataset. ( D ) Cell viability assay in different endocrine-sensitive or endocrine-resistant breast cancer cell lines (MCF7 Parental, MCF7- ESR1 Y537S , MCF7- ESR1 D538G , T47D Parental, T47D- ESR1 Y537S , T47D- ESR1 D538G , MCF7 cells treated with different treatments and BT474), cotreated with 4-OHT [10 −6 M], Fulv [10 −6 M] and Palb [10 −6 M] and SEL [10 −7 M]. A one-way analysis of variance (ANOVA) model was used for statistical significance of change in cell viability with different treatments. All values were presented as mean ± SEM from six independent repeats. ( E ) Scanning electron microscopy (SEM) images of human tissues and decellularized tissue from which ECM hydrogels are derived. ( F ) Stiffness of TissueSpec ® ECM Hydrogels. ( G ). MCF7- ESR1 Y537S cells were cultured at a density of 2 × 10 3 cells/well and embedded in 3D hydrogels to mimic different metastatic niches (bone, liver, lung) for breast cancer cells, and they were treated with different endocrine agents (4OHT [10 −6 ], Fulv [10 −6 ] and Palb [10 −6 ]) and in combination with SEL [10 −7 ]. ( H ). Colony size (pixels) and number were quantified using OpenCFU colony counting software ( http://opencfu.sourceforge.net/ ). A one-way analysis of variance (ANOVA) model was used for statistical significance of treatment and values were presented as mean ± SEM from three biological replicates (** p <0.01, *** p <0.001; **** p < 0.0001).

Article Snippet: IN SITE TM Metastasis Kit (Xylyx Bio, Inc., NY, USA) containing TissueSpec ® Bone (MTSBN101), Liver (MTSLV101) and Lung (MTSLG101) ECM Hydrogels), was used to model tumor microenvironments according to manufacturer protocol.

Techniques: Gene Expression, Expressing, RNA Sequencing, Viability Assay, Electron Microscopy, Derivative Assay, Cell Culture, Software