her2 positive cell line skbr3  (ATCC)

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: In silico analysis showing differential expression of glycogenes in the progression of BC: A Gene Set Enrichment Analysis (GSEA) showing the heatmap for different glycogenes in the HER2 high vs low phenotype. B Pie chart made using PANTHER software representing: cellular pathways of genes overexpressed in glycosylation geneset, and biological pathways of genes overexpressed in glycosylation geneset, C Cellular pathways of genes overexpressed in KRAS geneset and biological pathways of genes overexpressed in KRAS geneset. Major pathways highlighted in these pie charts have been indicated

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: In Silico, Quantitative Proteomics, Software, Glycoproteomics

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: In vitro analysis of ST3GAL1’s association with HER2: A Bar plot showing qRT-PCR analysis of ST3GAL1 in HER2 -ve and HER2 +ve BC cells. T-test based p-value obtained were as follows: SKBR3 vs MCF7 (p=0.0045, n=3), SKBR3 vs MDAMB231 (p=0.0034, n=3) and SKBR3 vs MDAMB435 (p=0.05, n=3). B Representative images of immunohistochemical analysis for ST3GAL1 in Grade 2 and 3 of Stage I, II and III BC tissues ( C ) ST3GAL1 expression in HER2 positive BC tissue. D Mean intensity value calculated as average pixel intensity using ImageJ, for ST3GAL1. Two-way ANOVA p-value exhibited p<0.05 (n=3) for Grade 2, Stage I vs Stage II vs Stage III. Similarly, p <0.05 (n=3) was obtained for Grade 3, Stage I vs Stage II vs Stage III. Moreover, t-test based p<0.05 (n=3) was obtained in the HER2 positive and HER2 negative BC tissues

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: In Vitro, Quantitative RT-PCR, Immunohistochemical staining, Expressing

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: In silico and in vitro analyses showing inverse association of GCNT3 with HER2: A Survival plot representing the gene expression of GCNT3 in BC cohorts. B Differential expression analysis of GCNT3 using the RNAseq data for the tumor vs control BC. C Differential expression analysis for the control vs tumor samples from the TCGA BC dataset for GCNT3 using GEPIA. D Bar plot showing qRT-PCR analysis for GCNT3 using the HER2 -ve and HER2 +ve BC cells with T-test based p-value obtained as follows: SKBR3 vs MCF7 (p=0.045, n=3), SKBR3 vs MDAMB231 (p=0.012, n=3) and SKBR3 vs MDAMB435 (p=0.00069, n=3). E Representative images of immunohistochemical analysis for GCNT3 in Grade 2 and 3 of Stage I, II and III BC tissues. F , G Mean intensity values calculated as average pixel intensity using ImageJ, for GCNT3. Two-way ANOVA p-value exhibited p<0.05 for Grade 2, Stage I vs Stage II vs Stage III. Similarly, p<0.05 (n=3) was obtained for Grade 3, Stage I vs Stage II vs Stage III. Additionally, t-test based p<0.05 (n=3) was obtained in the HER2 negative vs positive BC tissues

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: In Silico, In Vitro, Gene Expression, Quantitative Proteomics, RNA sequencing, Control, Quantitative RT-PCR, Immunohistochemical staining

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: In silico analysis of ST3GAL1’s association with HER2: A Survival plots representing gene expression of HER2 and ST3GAL1 for these genes in BC cohorts. B Oncomine extracted data represented by box plots on ST3GAL1 with increasing IHC score ( C ) Violin plots representing differential expression analysis of HER2 and ST3GAL for the tumor vs control using the RNAseq data for the BC samples. D Correlation plot between ST3GAL1 and HER2 on R2 dataset. E Differential expression analysis for the control vs tumor samples from the TCGA BC dataset for HER2 and ST3GAL1 using GEPIA. F Correlation plot between HER2 and ST3GAL1 on the cbioportal database

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: In Silico, Gene Expression, Quantitative Proteomics, Control, RNA sequencing

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: Functional analysis using GCNT3 inhibitor (talniflumate) in HER2 -ve and HER2 +ve BC cells: Representative images and quantification using talniflumate for ( A ) wound healing on MCF7 cells, t-test based p-value for control vs treatment at 24h and 48h were p=0.38 (n=3) and p=0.17 respectively, B colony formation on MCF7 cells, C wound healing on MDAMB231 cells, t-test based p-value for control vs treatment at 24h and 48h were p=0.023 (n=3) and p=0.084 (n=3) respectively, D colony formation on MDAMB231 cells, E wound healing on MDAMB435 cells, t-test based p-value for control vs treatment at 24h and 48h were p=0.09 (n=3) and p=0.40 (n=3) respectively, F colony formation on MDAMB435 cells, G . wound healing for SKBR3 cells, t-test based p-value for control vs treatment at 12h and 24h were p=0.49 (n=3) and p=0.32 (n=3) respectively and H colony formation for SKBR3 cells. NOTE: T-test based p-value calculation for colony formation assay were found to be p<0.05 (n=3) for all the cell lines

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: Functional Assay, Control, Colony Assay

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: Clinical significance of MUC1, β-catenin, Cyclin D1 in BC specimens: Representative images of immunohistochemical analysis in different stages and grades of BC for ( A ) β-catenin ( B ) MUC1 and C Cyclin D1. Mean intensity calculated of ( D ) β-catenin analyzed using two-way ANOVA p-value exhibited p<0.05 (n=3) for Grade 2, Stage I vs Stage II vs Stage III and Grade 3, Stage I vs Stage II vs Stage III, E MUC1 analyzed usingtwo-way ANOVA p-value exhibited p<0.05 (n=3) for Grade 2, Stage I vs Stage II, III and p>0.05 (n=3) for Grade 3, Stage I vs Stage II vs Stage III and F Cyclin D1 analyzed using two-way ANOVA p-value exhibited p<0.05 (n=3) for Grade 2, Stage I vs Stage II vs Stage III; however p>0.05 (n=3) was found for Grade 3 tissues. All the BC tissues herein were HER2 -ve

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: Immunohistochemical staining

Journal: BMC Cancer

Article Title: GCNT3 and ST3GAL1 expression correlates with HER2 status and MUC1/β-catenin/Cyclin D1 axis in breast cancer

doi: 10.1186/s12885-026-15821-w

Figure Lengend Snippet: Multiple analyses of GCNT3 in driving the MUC1/β-catenin/Cyclin D1 axis in BC: Representative immunohistochemistry images and mean intensity plots of HER2 +ve and HER2 -ve BC samples for (A ) β-catenin ( B ) MUC1 and C Cyclin D1. D Correlation plots of MUC1 with GCNT3, β-catenin, Cyclin D1 and ST3GAL1. E Gene interacting plot for GCNT3 with top 20 interacting nodes using STRING. F Scheme of the mechanistic link between GCNT3 and the β-catenin/Cyclin D1 axis in breast cancer. NOTE: T-test based p-value was found to be significant only for the Cyclin D1 stained HER2 positive vs negative BC tissues (p<0.05, n=3) whereas they were insignificant for the β-catenin and MUC1

Article Snippet: HER2 positive cell line SKBR3 (ATCC Catalog HTB-30) was cultured in McCoy’s 5 A medium.

Techniques: Immunohistochemistry, Staining

Journal: Thoracic Cancer

Article Title: Galectin‐3 enhances trastuzumab resistance by regulating cancer malignancy and stemness in HER2 ‐positive breast cancer cells

doi: 10.1111/1759-7714.14474

Figure Lengend Snippet: Galectin‐3 promoted cancer malignancy of HER2‐positive breast cancer cells. Western blot analysis of galectin‐3, HER2, and β‐actin in (a) pcDNA‐NC, pcDNA‐ LGALS3 , SKBR3, and SKBR3 cells treated with 100 ng/mL r‐Gal3 for 3 days; (b) JIMT1 cells transfected with siRNA‐NC, siRNA‐533, or siRNA‐571; and (c) JIMT1 cells treated with 0, 10, or 20 μg/mL GB1107 for 3 days. Cell viability curves for (d) pcDNA‐NC, pcDNA‐ LGALS3 , SKBR3, and SKBR3 cells treated with 100 ng/mL r‐Gal3 for 3 days; and (e) siRNA‐NC, siRNA‐533, siRNA‐571, and JIMT1 cells treated with 10 μg/mL GB1107 for 3 days. (f) Healing assays; (g) colony formation assays; and (h) transwell assays in pcDNA‐NC, pcDNA‐ LGALS3 , siRNA‐NC, and siRNA‐533 cells. The significant differences are indicated by asterisk (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001)

Article Snippet: The human HER2‐positive breast cancer cell lines SKBR3 (trastuzumab‐sensitive) and JIMT1 (trastuzumab‐resistant) were obtained from the American Type Culture Collection and maintained in McCoy's 5A medium and Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) (Gibco, Thermo Fisher Scientific), respectively.

Techniques: Western Blot, Transfection

Journal: Thoracic Cancer

Article Title: Galectin‐3 enhances trastuzumab resistance by regulating cancer malignancy and stemness in HER2 ‐positive breast cancer cells

doi: 10.1111/1759-7714.14474

Figure Lengend Snippet: Galectin‐3 activated Notch1 signaling pathway and promoted the cancer cell stemness of HER2‐positive breast cancer cells. (a) Immunofluorescence images of galectin‐3 (red) and DAPI (blue) in SKBR3‐CT cells, SKBR3‐OE cells, JIMT‐CT cells, and JIMT‐KO cells. (b) Western blot analysis of Notch1 pathway proteins (Notch1, NICD, HES1, and HEY1) and stemness biomarkers (CD24, CD44, CD133, Nanog, and E‐cadherin); β‐actin was used as a loading control. (c) Mammosphere formation in SKBR3‐OE, SKBR3‐CT, JIMT1‐KO, and JITM1‐CT cells. Representative images (top) and quantitative data (bottom) of mammosphere formation are shown. Scale bars, 100 μm. The significant differences are indicated by asterisk (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001)

Article Snippet: The human HER2‐positive breast cancer cell lines SKBR3 (trastuzumab‐sensitive) and JIMT1 (trastuzumab‐resistant) were obtained from the American Type Culture Collection and maintained in McCoy's 5A medium and Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) (Gibco, Thermo Fisher Scientific), respectively.

Techniques: Immunofluorescence, Western Blot, Control

Journal: Thoracic Cancer

Article Title: Galectin‐3 enhances trastuzumab resistance by regulating cancer malignancy and stemness in HER2 ‐positive breast cancer cells

doi: 10.1111/1759-7714.14474

Figure Lengend Snippet: Galectin‐3 affected the sensitivity of HER2‐positive breast cancer cells to trastuzumab. Cell viability curves and inhibition rate of (a) SKBR3‐CT and SKBR3‐OE cells and (b) JIMT1‐CT and JIMT1‐KO cells treated with trastuzumab for 72 hours. (c) Cell viability curves and inhibition rate of JIMT1‐CT cells treated with GB1107 for 72 hours. (d) Cell viability curves of JIMT1‐CT cells treated with GB1107 and/or trastuzumab. (e) Drug interaction analysis chart in JIMT1‐CT cells. The significant differences are indicated by asterisk (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001)

Article Snippet: The human HER2‐positive breast cancer cell lines SKBR3 (trastuzumab‐sensitive) and JIMT1 (trastuzumab‐resistant) were obtained from the American Type Culture Collection and maintained in McCoy's 5A medium and Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) (Gibco, Thermo Fisher Scientific), respectively.

Techniques: Inhibition

Journal: Nature Communications

Article Title: Disruption of FOXO3a-miRNA feedback inhibition of IGF2/IGF-1R/IRS1 signaling confers Herceptin resistance in HER2-positive breast cancer

doi: 10.1038/s41467-021-23052-9

Figure Lengend Snippet: a SKBR3, pool2, BT474, and HR20 cells cultured at normal condition were collected and subjected to western blot analyses of p-IGF-1R, IGF-1R, IRS1, p-Akt (T308) , p-Akt (S473) , Akt, p-S6K, S6K, p-FOXO3a, FOXO3a, and β-actin. b The levels of IGF1 and IGF2 in conditioned medium (CM) were measured by ELISA, **** p < 0.0001. c Pool2 or HR20 cells stably transfected with control shRNA (sh-Con) or IRS1 -targeting shRNAs (sh-1#, sh-2#) were treated with Herceptin at indicated concentrations for 72 h. Cell viability was evaluated by MTS assays, Pool2 sh-1#: ** p = 0.0021, **** p < 0.0001, sh-2#: ** p = 0.0096, **** p < 0.0001, HR20 sh-1#: * p = 0.0154, ** p = 0.0015, **** p < 0.0001, sh-2#: ** p = 0.0072, ** p = 0.0034, **** p < 0.0001. d Pool2 or HR20 cells with IRS1 deletion via CRISPR-Cas9 gene editing (sg-Con vs sg-1# and sg-2#) were treated with Herceptin at indicated concentrations for 72 h. Cell viability was evaluated by MTS assays, Pool2 sg-1#: *** p = 0.0002, **** p < 0.0001, sg-2#: * p = 0.0235, ** p = 0.0017, **** p < 0.0001, HR20 sg-1#: *** p = 0.0003, **** p < 0.0001, sg-2#: ** p = 0.0023, **** p < 0.0001. e Downregulation of IRS1 was achieved with either specific shRNAs (sh-Con vs sh-1# and sh-2#) or CRISPR-Cas9 gene editing (sg-Con vs sg-1# and sg-2#) in Pool2 or HR20 cells. The expression of IRS1, p-IGF-1R, IGF-1R, p-Akt (T308) , p-Akt (S473) , Akt, p-S6K, S6K, p-FOXO3a, FOXO3a, and β-actin was measured by western blot assays. f SKBR3 and BT474 cells were stably transfected with control vector (pLEX-Con or Con) or IRS1-overexpressing vector (pLEX-IRS1 or IRS1) followed by treatment with rhIGF2 (80 ng/ml) in combination with Herceptin at indicated concentrations for 72 h. Cell viability was evaluated by MTS assays, SKBR3 IRS1 + IGF2: ** p = 0.0059, *** p = 0.0002, *** p = 0.0003, BT474 IRS1 + IGF2: ** p = 0.0038, *** p = 0.0004. n = 3 biological independent samples ( b – d , f ). Data are presented as mean values ± SEM ( b – d , f ). Statistical significance was determined by a two-tailed Student’s t -test ( b – d , f ). Data show a representative of three independent experiments ( a , e ). All data are provided in the Source Data file.

Article Snippet: Human HER2-positive breast cancer cell lines SKBR3 and BT474 were obtained from the American Type Culture Collection (Manassas, VA, USA) and maintained in DMEM medium containing 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA).

Techniques: Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay, Stable Transfection, Transfection, Control, shRNA, CRISPR, Expressing, Plasmid Preparation, Two Tailed Test

Journal: Nature Communications

Article Title: Disruption of FOXO3a-miRNA feedback inhibition of IGF2/IGF-1R/IRS1 signaling confers Herceptin resistance in HER2-positive breast cancer

doi: 10.1038/s41467-021-23052-9

Figure Lengend Snippet: a SKBR3 cells were treated with rhIGF2 at indicated concentrations for 6 h. The expression of p-IGF-1R, IGF-IR, p-Akt (S473) , p-Akt (T308) , Akt, p-S6K, S6K, p-FOXO3a, FOXO3a, and β-actin was examined by western blot assays. b SKBR3 cells were transfected with a control empty vector (Con) or the same vector containing an IRS1 cDNA (IRS1), followed by rhIGF2 treatment for 6 h. The expression of indicated proteins was examined by western blot assays. c SKBR3 cells were transfected with control shRNA (sh-Con) or specific FOXO3a shRNAs (sh-1# and sh-2#) followed by rhIGF2 treatment for 6 h. The expression of indicated proteins was examined by western blot assays. d SKBR3 cells with IRS1 gene deletion via CRISPR-Cas9 (sg-Con vs sg-1# and sg-2#) were treated by rhIGF2 treatment for 6 h. The expression of indicated proteins were examined by western blot assays. Data show a representative of three independent experiments ( a – d ). All data are provided in the Source Data file.

Article Snippet: Human HER2-positive breast cancer cell lines SKBR3 and BT474 were obtained from the American Type Culture Collection (Manassas, VA, USA) and maintained in DMEM medium containing 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA).

Techniques: Expressing, Western Blot, Transfection, Control, Plasmid Preparation, shRNA, CRISPR

Journal: Nature Communications

Article Title: Disruption of FOXO3a-miRNA feedback inhibition of IGF2/IGF-1R/IRS1 signaling confers Herceptin resistance in HER2-positive breast cancer

doi: 10.1038/s41467-021-23052-9

Figure Lengend Snippet: a SKBR3 or BT474 cells were transfected with miR-128-3p or/and miR-30a-5p inhibitor (upper) or transfected with miR-128-3p or/and miR-30a-5p mimics (bottom). The expression of IRS1 and β-actin was examined by western blot assays. b SKBR3 or BT474 cells were transfected with miR-128-3p or/and miR-30a-5p inhibitor followed by rhIGF-2 treatment at indicated concentration for 24 h. The expression of IRS1 and β-actin was examined by western blot assays. c SKBR3 or BT474 cells were transfected with FOXO3a shRNA followed by rhIGF-2 treatment at indicated concentration for 24 h. The expression levels of miR-128-3p and miR-30a-5p were measured by qRT-PCR, **** p < 0.0001. d A schematic representation of FOXO3a binding sites within the 2 kb putative promoters of miR-128-3p and miR-30a-5p. The first base of the precursors of miR-128-3p and miR-30a-5p is defined as ‘+1’. e SKBR3 or BT474 cells were treated with rhIGF-2 at indicated concentrations for 24 h. The enrichment of FOXO3a at miR-128 or miR-30a promoter was evaluated by ChIP-qPCR. The chromatin was precipitated with an anti-FOXO3a antibody. The precipitated chromatin was then analyzed by qRT-PCR with primers specific for the putative FOXO3a binding sites, **** p < 0.0001. n = 3 biological independent samples ( c , e ). Data are presented as mean values ± SEM ( c , e ). Statistical significance was determined by a two-tailed Student’s t -test ( c , e ). Data show a representative of three independent experiments ( a , b ). All data are provided in the Source Data file.

Article Snippet: Human HER2-positive breast cancer cell lines SKBR3 and BT474 were obtained from the American Type Culture Collection (Manassas, VA, USA) and maintained in DMEM medium containing 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA).

Techniques: Transfection, Expressing, Western Blot, Concentration Assay, shRNA, Quantitative RT-PCR, Binding Assay, ChIP-qPCR, Two Tailed Test

Journal: Nature Communications

Article Title: Disruption of FOXO3a-miRNA feedback inhibition of IGF2/IGF-1R/IRS1 signaling confers Herceptin resistance in HER2-positive breast cancer

doi: 10.1038/s41467-021-23052-9

Figure Lengend Snippet: a, b SKBR3 or BT474 cells were treated with vehicle (Mock) or Rapamycin (10 μM, Rapa) in combination with rhIGF2 at indicated concentrations for 24 h. The expression of p-Akt (S473) , Akt, p-S6K, S6K, p-FOXO3a, FOXO3a, IRS1, PPP3CB, and β-actin was examined by western blot assays ( a ); The expression levels of miR-128-3p and miR-30a-5p were detected by qRT-PCR ( b ), **** p < 0.0001. c SKBR3 or BT474 cells were treated with a series of phosphatase inhibitors as Cantharidic acid (0.5 μM), Endothall (1 μM), RK-682 (10 μM), Cypermethrin (1 μM), Deltamethrin (1 μM), RWJ-60475 (2 μM), Tyrphostin 8 (10 μM), CinnGel (1 μM), BML-260 (10 μM), or BN-82002 (5 μM) in combination with rhIGF2 at indicated concentrations for 24 h. The expression of p-FOXO3a, FOXO3a, and β-actin was analyzed by western blot assays. d SKBR3 cells were treated with rhIGF2 at indicated concentrations for 24 h. The expression of PPP3CB, PPP3CA, PPP3CC, PPP3R1, PPP3R2, and β-actin was examined by western blot assays. e , f SKBR3 or BT474 cells with specific knockdown of PPP3CB by shRNAs (sh-Con vs sh-1# and sh-2#) were treated with rhIGF2 (80 ng/ml) for 24 h. The expression of PPP3CB, p-FOXO3a, FOXO3a and β-actin were examined by western blot assays ( e ); The enrichment of FOXO3a at the promoters of miR-128-3p and miR-30a-5p were detected by ChIP-qPCR ( f ), **** p < 0.0001. n = 3 biological independent samples ( b , f ). Data are presented as mean values ± SEM ( b , f ). Statistical significance was determined by a two-tailed Student’s t -test ( b , f ). Data show a representative of three independent experiments ( a , c , d , e ). All data are provided in the Source Data.

Article Snippet: Human HER2-positive breast cancer cell lines SKBR3 and BT474 were obtained from the American Type Culture Collection (Manassas, VA, USA) and maintained in DMEM medium containing 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Knockdown, ChIP-qPCR, Two Tailed Test

Journal: Nature Communications

Article Title: Disruption of FOXO3a-miRNA feedback inhibition of IGF2/IGF-1R/IRS1 signaling confers Herceptin resistance in HER2-positive breast cancer

doi: 10.1038/s41467-021-23052-9

Figure Lengend Snippet: a The expression levels of miR-128-3p and miR-30a-5p in SKBR3, pool2, BT474, or HR20 cells were measured by qRT-PCR, **** p < 0.0001. b Pool2 or HR20 cells were transfected with miR-128-3p or/and miR-30a-5p mimics. The expression of IRS1, p-FOXO3a, FOXO3a, and β-actin was examined by western blot assays. c Pool2 cells were transfected with miRNA mimics in combination with pLEX-IRS1 followed by Herceptin treatment at indicated concentrations for 72 h (left). SKBR3 cells were transfected miRNA inhibitors in combination with IRS1 shRNA, and then treated with 80 ng/ml rhIGF2 along with Herceptin (right). Cell viability was evaluated by MTS assays. Pool2 Mimics: ** p = 0.0022, *** p = 0.0003, **** p < 0.0001, Mimics+pLEX-IRS1: *** p = 0.0001, **** p < 0.0001, SKBR3 Inhibitors+IGF2: ** p = 0.0072, ** p = 0.0035, **** p < 0.0001, Inhibitors+IGF2 + sh-IRS1: ** p = 0.0079, *** p = 0.0002, **** p < 0.0001. d The expression levels of miR-193a-5p in SKBR3, pool2, BT474, and HR20 cells were detected by qRT-PCR, **** p < 0.0001. e Pool2 or HR20 cells were transfected with a miR-193a-5p mimic. SKBR3 or BT474 were transfected with a miR-193a-5p inhibitor. IGF2 levels in the CM were measured by ELISA. f Pool2 or HR20 cells were treated with vehicle (Mock) or WAY-600 (1 μM, WAY). The expression of p-Akt (S473) , Akt, p-S6K, S6K, p-FOXO3a, FOXO3a, and β-actin was examined by western blot assays (left); the expression levels of miR-128-3p, miR-30a-5p and miR-193a-5p were detected by qRT-PCR (right), **** p < 0.0001. g Pool2 or HR20 cells were treated with WAY-600 for 24 h. The enrichment of FOXO3a at miR-193a-5p promoter was examined by ChIP-qPCR assays, **** p < 0.0001. n = 3 biological independent samples ( a , c – e , g ). Data are presented as mean values ± SEM ( a , c – e , g ). Statistical significance was determined by a two-tailed Student’s t -test ( a , c – e , g ). Data show a representative of three independent experiments ( b , f ). All data are provided in the Source Data.

Article Snippet: Human HER2-positive breast cancer cell lines SKBR3 and BT474 were obtained from the American Type Culture Collection (Manassas, VA, USA) and maintained in DMEM medium containing 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, shRNA, Enzyme-linked Immunosorbent Assay, ChIP-qPCR, Two Tailed Test

Journal: Nature Communications

Article Title: Disruption of FOXO3a-miRNA feedback inhibition of IGF2/IGF-1R/IRS1 signaling confers Herceptin resistance in HER2-positive breast cancer

doi: 10.1038/s41467-021-23052-9

Figure Lengend Snippet: a The expression of PPP3CB, p-STAT6, STAT6, p-Src, Src, and β-actin in the indicated cells was analyzed by western blot assays (left). The expression levels of PPP3CB mRNA were measured by qRT-PCR (right), **** p < 0.0001. b Pool2 or HR20 cells with PPP3CB overexpression were treated with Herceptin for 72 h (top). SKBR3 or BT474 cells with specific knockdown of PPP3CB were treated with Herceptin for 72 h (bottom). Cell viability was evaluated by MTS assays, SKBR3 sh-1#: ** p = 0.008, ** p = 0.0013, *** p = 0.0002, sh-2#: ** p = 0.0019, **** p < 0.0001, BT474 sh-1#: ** p = 0.0038, *** p = 0.0002, *** p = 0.0001, sh-2#: *** p = 0.0003, *** p = 0.0001, **** p < 0.0001. c Pool2 or HR20 cells were transfected with control vector (Control) or PPP3CB-overexpressing vector (PPP3CB). The expression of PPP3CB, p-FOXO3a, FOXO3a, IRS1, and β-actin was examined by western blot assays (left). IGF2 levels in the CM were detected by ELISA (right). d The expression levels of miR-128-3p, miR-30a-5p, and miR-193a-5p in PPP3CB-overexpressing pool2 or HR20 cells were measured by qRT-PCR, **** p < 0.0001. e Pool2 or HR20 cells were treated with vehicle (Mock) or entinostat (1 μM, Ent.) for 48 h. The expression of PPP3CB, p-FOXO3a, FOXO3a, IRS1, and β-actin was examined by western blots. f , g Pool2 or HR20 cells were transfected with control shRNA (Con or sh-Con) or specific STAT6 shRNAs (sh-2# or sh-4#). The expression of STAT6, PPP3CB, p-FOXO3a, FOXO3a, IRS1, and β-actin was examined by western blots ( f ); the enrichment of HDAC1 at PPP3CB promoter was determined by ChIP-qPCR ( g ), **** p < 0.0001. h Total protein extracts of inducated cells were subjected to IP using an anti-HDAC1 antibody or control IgG, followed by western blot analysis of HDAC1 or p-STAT6. i Pool2 or HR20 cells treated with vehicle (Mock) or SU6656 (10 μM) were examined by western blot analysis. n = 3 biological independent samples ( a , b , c , d , g ). Data are presented as mean values ± SEM ( a , b , c , d , g ). Statistical significance was determined by a two-tailed Student’s t -test ( a , b , c , d , g ). All data are provided in the Source Data.

Article Snippet: Human HER2-positive breast cancer cell lines SKBR3 and BT474 were obtained from the American Type Culture Collection (Manassas, VA, USA) and maintained in DMEM medium containing 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Over Expression, Knockdown, Transfection, Control, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, shRNA, ChIP-qPCR, Two Tailed Test

Journal: Computational and Structural Biotechnology Journal

Article Title: Substantial cell apoptosis provoked by naked PAMAM dendrimers in HER2-positive human breast cancer via JNK and ERK1/ERK2 signalling pathways

doi: 10.1016/j.csbj.2021.05.011

Figure Lengend Snippet: (A-L). Effect of different concentrations of (A and B) G 6 NH 2 PAMAMs, (C and D) G 4 NH 2 PAMAMs, (E and F) G 6 OH PAMAMs, (G and H) G 5.5 COOH PAMAMs, and (I and J) lapatinib on cell viability of SKBR3 and ZR75 cells after 48 h of treatment. Effect of G 6 NH 2 and G 4 NH 2 PAMAMs on cell viability of MCF10A cells (I and J) after 48 h of treatment. Data are presented as a percentage of treatment relative to the control (Mean ± SEM; n = 3). Statistical analysis was performed using one-way analysis of variance (ANOVA). Tukey’s post-hoc test was conducted to compare treatment groups and results were stated as *statistically significant when p < 0.05 compared to the control. * p < 0.05, **p < 0.01, and *** p < 0.001.

Article Snippet: HER2-positive breast cancer cell lines SKBR3 and ZR75 were purchased from the American Type Tissue Culture (ATCC) (Rockville, MD, USA).

Techniques: Control

Journal: Computational and Structural Biotechnology Journal

Article Title: Substantial cell apoptosis provoked by naked PAMAM dendrimers in HER2-positive human breast cancer via JNK and ERK1/ERK2 signalling pathways

doi: 10.1016/j.csbj.2021.05.011

Figure Lengend Snippet: (A and B). Time response to treatment with PAMAM dendrimers. Time response to PAMAM dendrimers was investigated in (A) SKBR3 and (B) ZR75 cells. Cells were treated with G 4 NH 2 (10 µM), G 6 NH 2 (10 µM), G 6 OH (100 µM) and G 5.5 COOH (100 µM). Cell viability was assessed after 48 h of treatment. Data are presented as a percentage of treatment relative to the control (Mean ± SEM; n = 3).

Article Snippet: HER2-positive breast cancer cell lines SKBR3 and ZR75 were purchased from the American Type Tissue Culture (ATCC) (Rockville, MD, USA).

Techniques: Control

Journal: Computational and Structural Biotechnology Journal

Article Title: Substantial cell apoptosis provoked by naked PAMAM dendrimers in HER2-positive human breast cancer via JNK and ERK1/ERK2 signalling pathways

doi: 10.1016/j.csbj.2021.05.011

Figure Lengend Snippet: Morphological changes induced by cationic PAMAM dendrimers and lapatinib. SKBR3, ZR75 and MCF10A cells were treated with 5 µM of G 4 NH 2 and G 6 NH 2 . Images were taken at a magnification scale of 10X following 48 h of treatment (n = 3). SKBR3 and ZR75 cells were treated with lapatinib and morphological images were taken at a magnification scale of 10X following 48 h of treatment (n = 3).

Article Snippet: HER2-positive breast cancer cell lines SKBR3 and ZR75 were purchased from the American Type Tissue Culture (ATCC) (Rockville, MD, USA).

Techniques:

Journal: Computational and Structural Biotechnology Journal

Article Title: Substantial cell apoptosis provoked by naked PAMAM dendrimers in HER2-positive human breast cancer via JNK and ERK1/ERK2 signalling pathways

doi: 10.1016/j.csbj.2021.05.011

Figure Lengend Snippet: (A -D). Induction of apoptosis by PAMAM dendrimers in (A and B) SKBR3 and (C and D) ZR75 cells as determined by Annexin V apoptosis assay. Cells were treated with 5 µM of G 4 NH 2 , G 6 NH 2 , G 6 OH and G 5.5 COOH PAMAMs. Statistical analysis was performed using one-way analysis of variance (ANOVA). Tukey’s post-hoc test was conducted to compare treatment groups and results were stated as *statistically significant when p < 0.05 compared to the control. * p < 0.05, **p < 0.01, and *** p < 0.001.

Article Snippet: HER2-positive breast cancer cell lines SKBR3 and ZR75 were purchased from the American Type Tissue Culture (ATCC) (Rockville, MD, USA).

Techniques: Apoptosis Assay, Control

Journal: Computational and Structural Biotechnology Journal

Article Title: Substantial cell apoptosis provoked by naked PAMAM dendrimers in HER2-positive human breast cancer via JNK and ERK1/ERK2 signalling pathways

doi: 10.1016/j.csbj.2021.05.011

Figure Lengend Snippet: (A and B). Effect of PAMAM dendrimers on colony formation, in soft agar, in HER2-positive breast cancer cell lines, (A) SKBR3 and (B) ZR75. Cells were treated with 5 µM of G 4 NH 2 , G 6 NH 2 , G 6 OH and G 5.5 COOH PAMAMs. PAMAM dendrimers inhibit colony formation of SKBR3 and ZR75 in comparison with their matched control cells. Colonies were counted manually and expressed as a percentage of treatment relative to the control (Mean ± SEM). Statistical analysis was performed using one-way analysis of variance (ANOVA). Tukey’s post-hoc test was conducted to compare treatment groups and results were stated as *statistically significant when p < 0.05 compared to the control. * p < 0.05, **p < 0.01, and *** p < 0.001.

Article Snippet: HER2-positive breast cancer cell lines SKBR3 and ZR75 were purchased from the American Type Tissue Culture (ATCC) (Rockville, MD, USA).

Techniques: Comparison, Control

Journal: Computational and Structural Biotechnology Journal

Article Title: Substantial cell apoptosis provoked by naked PAMAM dendrimers in HER2-positive human breast cancer via JNK and ERK1/ERK2 signalling pathways

doi: 10.1016/j.csbj.2021.05.011

Figure Lengend Snippet: (A and B). RNA expression and molecular mechanisms of PAMAM dendrimers inhibitory actions in (A) SKBR3 and (B) ZR75 cell lines. PAMAMs induce deregulation of pro-apoptotic markers (Bax, Caspases-3, −8 and −9) in comparison with their control and inhibit anti-apoptotic markers (Bcl-2). Cells were treated with: G 4 NH 2 and G 6 NH 2 PAMAMs and lapatinib. GAPDH was used as a control for gene expression in this assay. Data are presented as a percentage of treatment relative to the control (Mean ± SEM; n = 3). Statistical analysis was performed using one-way analysis of variance (ANOVA). Tukey’s post-hoc test was conducted to compare treatment groups and results were stated as *statistically significant when p < 0.05 compared to the control. * p < 0.05, **p < 0.01, and *** p < 0.001.

Article Snippet: HER2-positive breast cancer cell lines SKBR3 and ZR75 were purchased from the American Type Tissue Culture (ATCC) (Rockville, MD, USA).

Techniques: RNA Expression, Comparison, Control, Gene Expression

Journal: Computational and Structural Biotechnology Journal

Article Title: Substantial cell apoptosis provoked by naked PAMAM dendrimers in HER2-positive human breast cancer via JNK and ERK1/ERK2 signalling pathways

doi: 10.1016/j.csbj.2021.05.011

Figure Lengend Snippet: (A and B). Protein expression and molecular mechanisms of PAMAM dendrimers inhibitory actions in (A) SKBR3 and (B) ZR75 cell lines. Cells were treated with: G 4 NH 2 , G 6 NH 2 PAMAMs, and lapatinib. GAPDH was used as a control for the loaded amount of the protein in this assay. Data are presented as a percentage of treatment relative to the control (Mean ± SEM; n = 3). Statistical analysis was performed using one-way analysis of variance (ANOVA). Tukey’s post-hoc test was conducted to compare treatment groups and results were stated as *statistically significant when p < 0.05 compared to the control. * p < 0.05, **p < 0.01, and *** p < 0.001.

Article Snippet: HER2-positive breast cancer cell lines SKBR3 and ZR75 were purchased from the American Type Tissue Culture (ATCC) (Rockville, MD, USA).

Techniques: Expressing, Control