Journal: bioRxiv

Article Title: The role of heparan sulfate in enhancing the chemotherapeutic response in triple-negative breast cancer

doi: 10.1101/2023.09.08.556819

Figure Lengend Snippet: (A) Basal heparan sulfate expression was examined in the supernatants of TNBC cells and control immortal MCF-10A via ELISA analysis with MDA-MB 468 expressing the most. The standard deviation was calculated from three independent experiments performed in triplicate. The student’s t-test was performed to determine the significance with * representing p<0.05 and ** representing p<0.01 the protein expression in MCF-10A to the protein expressions in the TNBC cell lines. (B) Baseline heparan sulfate cell surface expression levels were examined via flow cytometry analysis in nontumorigenic immortal mammary epithelial MCF-10A cells and TNBC MDA-MB 231, Hs 578t and MDA-MB 468 cells. The standard deviation was calculated from three independent experiments performed in triplicate. The student’s t-test was performed with * representing p<0.05 and ** representing p<0.01 comparing expression levels in MCF-10A to those in the TNBC cell lines. (C) The AMSBIO BR1202B breast cancer tissue array (120 cores with 82 TNBC cores; key can be found in Supplemental Figure 8C and corresponding Supplemental Table 1 showing breast cancer sub-type distribution) was stained with heparan sulfate.

Article Snippet: An AMSBIO breast cancer tissue array (120 cores specifically with 82 TNBC cores), two normal breast tissue slides and three DCIS tissue slides were stained with heparan sulfate specific antibody ( , Supplemental Figure 8) and statistical analysis was performed ( ).

Techniques: Expressing, Control, Enzyme-linked Immunosorbent Assay, Standard Deviation, Flow Cytometry, Staining

Journal: bioRxiv

Article Title: The role of heparan sulfate in enhancing the chemotherapeutic response in triple-negative breast cancer

doi: 10.1101/2023.09.08.556819

Figure Lengend Snippet: (A) Pairwise comparisons using Dunn’s test indicated that there was a significant difference between TNBC and normal breast (p = 0.0040), TNBC and DCIS (p = 0.0359), ER+/PR+ and normal (p = 0.0209), and HER2+ and normal (p = 0.0403), in the percentage of heparan sulfate positively stained cells in the tissue sections. No other differences were significantly different. (B) Pairwise comparisons using Dunn’s test showed that there was a significant difference between TNBC and normal (p = 0.0048), TNBC and DCIS (p = 0.0416), ER+/PR+ and normal (p = 0.0131), and HER2+ and normal (p = 0.0321), in the percentage of heparan sulfate weakly stained cells. No other differences were significantly different. (C) Pairwise comparisons using Dunn’s test demonstrated that there was a significant difference between TNBC and normal (p = 0.0003), TNBC and DCIS (p = 0.0064), ER+/PR+ and normal (p = 0.0127), HER2+ and normal (p = 0.0041), and HER2+ and DCIS (p = 0.0305), in the percentage of heparan sulfate moderately stained cells. No other differences were significantly different. (D) Pairwise comparisons using Dunn’s test demonstrated that there was a significant difference between TNBC and Normal (p = 0.0001), TNBC and DCIS (p = 0.0020), ER+/PR+ and normal (p = 0.0002), ER+/PR+ and DCIS (p = 0.0087), HER2+ and normal (p = 0.0003), and HER2+ and DCIS (p = 0.0109), in the percentage of heparan sulfate strongly stained cells. No other differences were significantly different. (E) Pairwise comparisons using Dunn’s test demonstrated that there was a significant difference between TNBC and normal (p = 0.0040), TNBC and DCIS (p = 0.0359), ER+/PR+ and normal (p = 0.0209), HER2+ and normal (p = 0.0003), and HER2+ and normal (p = 0.0403), in the percentage of cells negative for heparan sulfate staining. No other differences were significantly different. (F) Kruskal-Wallis test indicated that there was no significant difference in the percentage of heparan sulfate positively stained cells in the tumor amongst the TNBC breast cancer stages.

Article Snippet: An AMSBIO breast cancer tissue array (120 cores specifically with 82 TNBC cores), two normal breast tissue slides and three DCIS tissue slides were stained with heparan sulfate specific antibody ( , Supplemental Figure 8) and statistical analysis was performed ( ).

Techniques: Staining

Journal: bioRxiv

Article Title: The role of heparan sulfate in enhancing the chemotherapeutic response in triple-negative breast cancer

doi: 10.1101/2023.09.08.556819

Figure Lengend Snippet: (A) Baseline extracellular heparanase expression was determined in nontumorigenic immortal mammary epithelial MCF-10A cells and TNBC MDA-MB 231, Hs 578t and MDA-MB 468 cells by ELISAs. The standard deviation was calculated from three independent experiments performed in triplicate. The student’s t-test was performed with * representing p<0.05 and ** representing p<0.01, comparing expression levels in MCF-10A to those in the TNBC cell lines. (B) The AMSBIO BR1202B breast cancer tissue array (120 cores with 82 TNBC cores; key can be found in Supplemental Figure 8C and corresponding Excel worksheet showing breast cancer sub-type distribution) was stained with heparanase.

Article Snippet: An AMSBIO breast cancer tissue array (120 cores specifically with 82 TNBC cores), two normal breast tissue slides and three DCIS tissue slides were stained with heparan sulfate specific antibody ( , Supplemental Figure 8) and statistical analysis was performed ( ).

Techniques: Expressing, Standard Deviation, Staining

Journal: bioRxiv

Article Title: The role of heparan sulfate in enhancing the chemotherapeutic response in triple-negative breast cancer

doi: 10.1101/2023.09.08.556819

Figure Lengend Snippet: Kruskal-Wallis test indicated that there was no significant difference: (A) in the percentage of heparanase positively stained cells in the tissue sections of normal breast tissue, DCIS and invasive breast cancer subtypes, (B) in the percentage of heparanase weakly stained cells in the tissue sections of normal breast tissue, DCIS and invasive breast cancer subtypes, (C) in the percentage of heparanase moderately stained cells in the tissue sections of normal breast tissue, DCIS and invasive breast cancer subtypes, (D) in the percentage of heparanase strongly stained cells in the tissue sections of normal breast tissue, DCIS and invasive breast cancer subtypes, (E) in the percentage of negative heparanase stained cells in the tissue sections of normal breast tissue, DCIS and invasive breast cancer subtypes and (F) in the percentage of heparanase positively stained cells in the tissue sections of different TNBC breast cancer stages.

Article Snippet: An AMSBIO breast cancer tissue array (120 cores specifically with 82 TNBC cores), two normal breast tissue slides and three DCIS tissue slides were stained with heparan sulfate specific antibody ( , Supplemental Figure 8) and statistical analysis was performed ( ).

Techniques: Staining

Journal: Cancer Biology & Therapy

Article Title: Sulfatase 2 inhibition sensitizes triple-negative breast cancer cells to paclitaxel through augmentation of extracellular ATP

doi: 10.1080/15384047.2025.2483989

Figure Lengend Snippet: ELISA analysis of sulfatases and SULF2 immunohistochemistry. ELISAs were performed to examine the basal levels of (a) SULF1 and (b) SULF2 in adjusted cell supernatants inversely proportionate to the protein concentration in the corresponding cell lysate of MCF-10A cell line, TNBC MDA-MB 231, Hs 578t and MDA-MB 468 cell lines and ER+/PR+ T47D, MCF-7 and ZR-75-1 cell lines. MCF-10A expressed more SULF1 in comparison to the TNBCs but similar expression levels were observed in ER+/PR+ T47D and MCF-7 cells. However, the ER+/PR+ ZR-75-1 cell line expressed the most SULF1. Meanwhile, TNBCs and ER+/PR+ T47D and ZR-75-1 cells expressed more SULF2 in comparison to MCF-10A cells. Standard deviation was calculated from three independent experiments performed in triplicate. The student’s t-test was performed to determine significance with *representing p < 0.05 and **representing p < 0.01 comparing the protein expression MCF-10A to the protein expressions of TNBC cell lines. (b) The AMSBIO BR1202B breast cancer tissue array (120 cores with 82 TNBC cores, 20 ER+/PR+ cores, 14 HER2+ cores, and 4 necrotic cores; key can be found in supplemental Figure S6C) was stained with SULF2.

Article Snippet: The student’s t-test was performed to determine significance with *representing p < 0.05 and **representing p < 0.01 comparing the protein expression MCF-10A to the protein expressions of TNBC cell lines. (b) The AMSBIO BR1202B breast cancer tissue array (120 cores with 82 TNBC cores, 20 ER+/PR+ cores, 14 HER2+ cores, and 4 necrotic cores; key can be found in supplemental Figure S6C) was stained with SULF2.

Techniques: Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Protein Concentration, Comparison, Expressing, Standard Deviation, Staining

Journal: Cancer Biology & Therapy

Article Title: Sulfatase 2 inhibition sensitizes triple-negative breast cancer cells to paclitaxel through augmentation of extracellular ATP

doi: 10.1080/15384047.2025.2483989

Figure Lengend Snippet: Statistical analysis for SULF2 immunohistochemistry. For the breast cancer tissue array and slides stained for heparanase: TNBC ( n =75), ER+/PR+ ( n =18), HER2+ ( n =14), (a) images were taken of SULF2-stained AMSBIO BR1202B breast cancer tissue array on an evos FL auto 2 microscope (40×). (b) Pairwise comparisons using Dunn’s test indicated that there was a significant difference in H-score between breast cancer sub-types TNBC and ER+/PR+ ( p = 0.0392). There was no significant different between TNBC and HER2+. (c) The Kruskal-Wallis test indicated that there was no significant difference in the average percentages of cells that stained positively for SULF2 in tissue sections of TNBC, ER+/PR+ breast cancer, and HER2+ breast cancer. (d) Pairwise comparisons using Dunn’s test indicated that there was a significant difference between breast cancer sub-types TNBC and ER+/PR+ ( p = 0.009), and TNBC and HER2+ ( p = 0.0338), in the percentages of cells that stained moderately or strongly positive for SULF2. (e) The same Dunn’s test from (d) also indicated that there was a significant difference between cancer sub-types TNBC and ER+/PR+ ( p = 0.0094), and TNBC and HER2+( p = 0.0338), in the percentages of cells that stained negative or weakly positive for SULF2. (f) Pairwise comparisons using Dunn’s test showed that there was a significant difference between TNBC breast cancer stages 2A and 2B ( p = 0.0007) in the percentages of cells that stained positively for SULF2. No other differences among different TNBC stages were significant. Standard error of the mean was calculated. For statistical analysis, *representing p < 0.05, **representing p < 0.01 and ***representing p < 0.001.

Article Snippet: The student’s t-test was performed to determine significance with *representing p < 0.05 and **representing p < 0.01 comparing the protein expression MCF-10A to the protein expressions of TNBC cell lines. (b) The AMSBIO BR1202B breast cancer tissue array (120 cores with 82 TNBC cores, 20 ER+/PR+ cores, 14 HER2+ cores, and 4 necrotic cores; key can be found in supplemental Figure S6C) was stained with SULF2.

Techniques: Immunohistochemistry, Staining, Microscopy