fluorescent in situ hybridization (fish) utilizing pathvision her2 dna kit  (Abbott Laboratories)

Journal: NPJ Precision Oncology

Article Title: Real life outcome analysis of breast cancer brain metastases treated with Trastuzumab Deruxtecan

doi: 10.1038/s41698-025-00801-3

Figure Lengend Snippet: Baseline characteristics

Article Snippet: Eligible patients were required to have a diagnosis of HER2-positive tumor, determined locally and defined as 3+ immunohistochemical (IHC) staining (HercepTest; Dako A/S, Glostrup, Denmark) or 2 + IHC staining and HER2/Vep17 Ratio > 2 at fluorescence in situ hybridization (FISH) (PathVision HER2 DNA probe kit; Vysis Inc., Downers Grove, IL).

Techniques: Immunohistochemistry, Amplification, Adjuvant

Journal: Cancers

Article Title: HER2 mRNA Levels, Estrogen Receptor Activity and Susceptibility to Trastuzumab in Primary Breast Cancer

doi: 10.3390/cancers14225650

Figure Lengend Snippet: Association between HER2 mRNA expression, HER2-addiction and patient prognosis. ( a , b ) Association between PAM50 ( a ) and TRAR ( b ) classification and HER2 mRNA expression in primary HER2-positive BCs of the GHEA cohort. p values obtained by Student’s t test. ( c – e ) Kaplan-Meier analysis of patients in the GHEA cohort classified according to HER2 mRNA levels in the entire cohort (n = 36, c ), and in ER-negative (n = 18, d ) and ER-positive (n = 18, e ) cohorts. The HER2 mRNA median expression value in the entire cohort was used as the cutoff to define the high and low expression groups. p values by log-rank test. HER2-E: HER2-enriched.

Article Snippet: The status of HER2/neu was investigated utilizing a commercial probe PathVision HER2/neu DNA probe kit (Vysis, Abbot Molecular, Des Plaines, IL, USA) on metaphase spreads obtained from BC cell lines using standard cytogenetic methodologies [ ].

Techniques: Expressing

Journal: Cancers

Article Title: HER2 mRNA Levels, Estrogen Receptor Activity and Susceptibility to Trastuzumab in Primary Breast Cancer

doi: 10.3390/cancers14225650

Figure Lengend Snippet: Association between HER2 protein expression and patient prognosis. ( a ) Representative images of HER2 positivity (red) by IF are shown. Images were analyzed using the same parameters. Tumors were scored as 3+ (1st tertile), 4+ (2nd tertile) and 5+ (3rd tertile) according to the mean fluorescence intensities of positive pixels. Nuclei were stained with DAPI (blue). ( b , c ) Association between HER2 mRNA expression levels (log2) and protein levels determined according to HER2 IF-score (log2) in ER-negative ( b ) and ER-positive ( c ) subgroups. The Pearson coefficient and relative p value are shown. ( d , e ) Kaplan-Meier analysis of ER-negative (n = 17, d ) and ER-positive (n = 16, e ) patients classified according to the HER2 IF-score. p value determined by log-rank test.

Article Snippet: The status of HER2/neu was investigated utilizing a commercial probe PathVision HER2/neu DNA probe kit (Vysis, Abbot Molecular, Des Plaines, IL, USA) on metaphase spreads obtained from BC cell lines using standard cytogenetic methodologies [ ].

Techniques: Expressing, Fluorescence, Staining

Journal: Cancers

Article Title: HER2 mRNA Levels, Estrogen Receptor Activity and Susceptibility to Trastuzumab in Primary Breast Cancer

doi: 10.3390/cancers14225650

Figure Lengend Snippet: Correlation between HER2 mRNA and tumor molecular characteristics according to ER expression. ( a , b ) Bar plot showing HALLMARK pathways significantly (FDR < 10%) and positively (red bars) or negatively (green bars) correlated with HER2 mRNA in ER-negative ( a ) and ER-positive ( b ) HER2+ BCs of the GHEA cohort as evaluated by GSEA. FDR: false discovery rate. Gray arrows: proliferation-related pathways; red arrows: oncogene downstream signaling pathways; black arrows: immune-related pathways. ( c ) Proteins significantly ( p < 0.05) correlated with HER2 mRNA levels in the TCGA dataset. Pearson’s r is color-co3.4. Association between HER2 Expression and HER2-Addiction in BC Cell Lines.

Article Snippet: The status of HER2/neu was investigated utilizing a commercial probe PathVision HER2/neu DNA probe kit (Vysis, Abbot Molecular, Des Plaines, IL, USA) on metaphase spreads obtained from BC cell lines using standard cytogenetic methodologies [ ].

Techniques: Expressing

Journal: Cancers

Article Title: HER2 mRNA Levels, Estrogen Receptor Activity and Susceptibility to Trastuzumab in Primary Breast Cancer

doi: 10.3390/cancers14225650

Figure Lengend Snippet: Characterization of HER2-positive BC cell lines. ( a ) Lapatinib dose–response curves for HER2-positive BC cell lines. Results are normalized to those of untreated cells and are representative of two independent experiments. ( b ) Western blot analysis of HER2-positive BC cell extracts after immunoprecipitation with anti-HER2 antibodies. Images are representative of two independent experiments. Ratios were calculated for band quantification by Quantity One. ER-positive and HER2-addicted BC cell lines are shown in red and underlined, respectively. ( c ) HER2 mRNA levels as evaluated by qRT-PCR. Data are the mean ± SD of three independent experiments and are relative to BT474 HER2 expression levels. ( d ) Western blot analysis of HER2-positive BC cell extracts. Vinculin was used as a loading control. Images are representative of two independent experiments. The names of ER-positive and HER2-addicted BC cell lines are shown in red and underlined, respectively. The asterisks indicate molecules analyzed in a separate gel.

Article Snippet: The status of HER2/neu was investigated utilizing a commercial probe PathVision HER2/neu DNA probe kit (Vysis, Abbot Molecular, Des Plaines, IL, USA) on metaphase spreads obtained from BC cell lines using standard cytogenetic methodologies [ ].

Techniques: Western Blot, Immunoprecipitation, Quantitative RT-PCR, Expressing

Journal: Cancers

Article Title: HER2 mRNA Levels, Estrogen Receptor Activity and Susceptibility to Trastuzumab in Primary Breast Cancer

doi: 10.3390/cancers14225650

Figure Lengend Snippet: Modulation of ligand-dependent ER activity in ER-positive, HER2-positive BC cell lines. ( a ) HER2 and PGR mRNA levels, as evaluated by qRT-PCR in cells grown in complete medium and treated with 10 nM fulvestrant for 24 h. Data are relative to cells treated with diluent (dotted line), are the mean ± SD of technical replicates and are representative of 3 independent experiments. ( b ) HER2 mRNA levels as evaluated by qRT-PCR in cells grown in complete medium containing 10% FBS (with phenol red) or in white medium with 10% charcoal-stripped FBS, treated with 10 nM estradiol (white + E2) or not (white) for 24 h. Data are the mean ± SD of technical replicates and are representative of 3 independent experiments. ( c ) PGR mRNA expression in cells treated as in ( b ). Data are the mean ± SD of technical replicates and are representative of 3 independent experiments. ( d ) Western blot analysis of cells treated as in b. Representative of two independent experiments. Vinculin was used as a loading control. The asterisks indicate molecules analyzed in a separate gel. * p < 0.05 by unpaired Student’s t test. RQ: relative quantification.

Article Snippet: The status of HER2/neu was investigated utilizing a commercial probe PathVision HER2/neu DNA probe kit (Vysis, Abbot Molecular, Des Plaines, IL, USA) on metaphase spreads obtained from BC cell lines using standard cytogenetic methodologies [ ].

Techniques: Activity Assay, Quantitative RT-PCR, Expressing, Western Blot

Journal: Cancers

Article Title: HER2 mRNA Levels, Estrogen Receptor Activity and Susceptibility to Trastuzumab in Primary Breast Cancer

doi: 10.3390/cancers14225650

Figure Lengend Snippet: Correlation between E2 levels and tumor characteristics in BC patients. ( a , b ) Bar plot showing HALLMARK pathways significantly (FDR < 15%) positively (red bars) or negatively (green bars) correlated with E2 (pg/mL quantified in plasma of patients at surgery) in 10 postmenopausal HER2-positive BC patients of the GHEA cohort ( a ) and in the ER-positive subgroup (n = 5, b ) as evaluated by GSEA. FDR: false discovery rate. ( c ) Correlation of E2 with the expression of known ER-regulated genes in the same cohort analyzed in a. Pearson’s r is color-coded, and its significance is shown: # p < 0.1, * p < 0.05, ** p < 0.01. ( d , e ) Kaplan-Meier analysis of postmenopausal patients with HER2-positive BC treated with adjuvant trastuzumab classified according to circulating E2 levels in the entire (E2 high n = 20, E2 low: n = 20, d ) and ER-positive (E2 high n = 10, E2 low: n = 13, e ) cohorts. The E2 median value in the entire cohort was used as a cutoff to define the high- and low-level groups. p values by log-rank test.

Article Snippet: The status of HER2/neu was investigated utilizing a commercial probe PathVision HER2/neu DNA probe kit (Vysis, Abbot Molecular, Des Plaines, IL, USA) on metaphase spreads obtained from BC cell lines using standard cytogenetic methodologies [ ].

Techniques: Expressing