non ibc cell line skbr3  (ATCC)

Journal: Molecular Cancer

Article Title: Syndecan-1 is a novel molecular marker for triple negative inflammatory breast cancer and modulates the cancer stem cell phenotype via the IL-6/STAT3, Notch and EGFR signaling pathways

doi: 10.1186/s12943-017-0621-z

Figure Lengend Snippet: Expression of Syndecan-1 and the CSC marker CD44 in carcinoma tissues of IBC vs non-IBC patients, SUM-149 and SKBR3 cells. a Higher expression of Syndecan-1 mRNA level in carcinoma tissues of IBC ( n = 13) vs non-IBC ( n = 14). RQ values of mRNA expression are log2-transformed and normalized to values of normal tissues collected during reduction mammoplasty. Bars represent median with interquartile range. ** P < 0.01 as determined by Mann-Whitney U-test. b Representative fields of immunostaining ( brown color ) of Syndecan-1 and CD44 in paraffin embedded carcinoma tissue sections of triple negative IBC ( n = 13) and non-IBC ( n = 17) patients. A high density of cancer cells positive for CD44 and Syndecan-1 is observed in IBC vs non-IBC. c Pearson’s correlation between Syndecan-1 and CD44 expression in carcinoma tissues of IBC vs non-IBC. d A representative flow cytometric analysis for the expression of CD44 and Syndecan-1 in SUM-149 and SKBR3 cells. e Quantitative analysis of four subpopulations; CD44 (-) Syndecan-1 (-) , CD44 (+) Syndecan-1 (-) , CD44 (-) Syndecan-1 (+) and CD44 (+) Syndecan-1 (+) . Syndecan-1 is higher expressed in the CD44 (+) -enriched subset in SUM-149 cells than that in SKBR3 cells. Data represent mean ± SEM, n ≥ 3. ** P < 0.01, # P < 0.001 as determined by Student’s t-test. Data shown are a single experiment representative of three independent experiments

Article Snippet: The human IBC cell line SUM-149 (a kind gift from Dr. Bonnie Sloane, Wayne State University, Detroit, MI, USA) and the non-IBC cell line SKBR3 (ATCC/LGC Promochem, Wesel, Germany) were maintained in HAM’s-F12 and DMEM containing 10% FCS, 1% glutamine and 1% penicillin/streptomycin in a humidified atmosphere of 5% CO 2 at 37 °C, respectively.

Techniques: Expressing, Marker, Transformation Assay, MANN-WHITNEY, Immunostaining

Journal: Molecular Cancer

Article Title: Syndecan-1 is a novel molecular marker for triple negative inflammatory breast cancer and modulates the cancer stem cell phenotype via the IL-6/STAT3, Notch and EGFR signaling pathways

doi: 10.1186/s12943-017-0621-z

Figure Lengend Snippet: Syndecan-1 silencing suppresses CSC-related gene expression in SUM-149 and SKBR3 cells. Post Syndecan-1 knockdown, total RNA isolated from SUM-149 ( a ) and SKBR3 cells ( b ) was reverse transcribed into cDNA and subjected into qPCR. Data represent mean ± SEM, n ≥ 3. * P < 0.05, # P < 0.01 as determined by Student’s t-test

Article Snippet: The human IBC cell line SUM-149 (a kind gift from Dr. Bonnie Sloane, Wayne State University, Detroit, MI, USA) and the non-IBC cell line SKBR3 (ATCC/LGC Promochem, Wesel, Germany) were maintained in HAM’s-F12 and DMEM containing 10% FCS, 1% glutamine and 1% penicillin/streptomycin in a humidified atmosphere of 5% CO 2 at 37 °C, respectively.

Techniques: Gene Expression, Knockdown, Isolation, Reverse Transcription

Journal: Molecular Cancer

Article Title: Syndecan-1 is a novel molecular marker for triple negative inflammatory breast cancer and modulates the cancer stem cell phenotype via the IL-6/STAT3, Notch and EGFR signaling pathways

doi: 10.1186/s12943-017-0621-z

Figure Lengend Snippet: Syndecan-1 silencing attenuates the activation of STAT-3 and NFκB signaling pathways and downregulates protein expression of gp130 in SUM-149 and SKBR3 cells. Seventy-two hours post transfection total cell lysates of control and Syndecan-1 knockdown cells were collected, electrophoresed and immunoblotted. The membrane was probed with the indicated antibodies. a Western blot showing expression of p-STAT-3, STAT-3, p-NFκB, NFκB and gp130 upon Syndecan-1 silencing in SUM-149 and SKBR-3 cells. b Immunoblot band intensities were normalized to the total form of STAT-3, NFκB or tubulin expression. Data shown are a single experiment representative of three independent experiments. * P < 0.05, ** P < 0.01 and # P < 0.001 as determined by Student’s t-test

Article Snippet: The human IBC cell line SUM-149 (a kind gift from Dr. Bonnie Sloane, Wayne State University, Detroit, MI, USA) and the non-IBC cell line SKBR3 (ATCC/LGC Promochem, Wesel, Germany) were maintained in HAM’s-F12 and DMEM containing 10% FCS, 1% glutamine and 1% penicillin/streptomycin in a humidified atmosphere of 5% CO 2 at 37 °C, respectively.

Techniques: Activation Assay, Protein-Protein interactions, Expressing, Transfection, Control, Knockdown, Membrane, Western Blot

Journal: Molecular Cancer

Article Title: Syndecan-1 is a novel molecular marker for triple negative inflammatory breast cancer and modulates the cancer stem cell phenotype via the IL-6/STAT3, Notch and EGFR signaling pathways

doi: 10.1186/s12943-017-0621-z

Figure Lengend Snippet: Syndecan-1 is a modulator of EGFR expression and activation via Notch signaling in IBC. a Scatter plot shows a significant upregulation of EGFR mRNA in tissues of IBC ( n = 13) vs non-IBC ( n = 14). RQ values of EGFR mRNA expression in tissues of IBC vs non-IBC are log2-transformed and normalized to values of normal tissues collected during reduction mammoplasty. Bars represent median with interquartile range. * P < 0.05 as determined by Mann-Whitney U-test. b Pearson’s correlation between Syndecan-1 and EGFR mRNA expression in tissues of non-IBC tissues ( left panel ) and IBC ( right panel ). c EGFR mRNA and protein level expression in control and Syndecan-1-silenced SUM-149 and SKBR3 cells. ** P < 0.01 and # P < 0.001 as determined by Student’s t-test. d Expression of EGFR mRNA levels in control and Syndecan-1-silenced SUM-149 cells post GSI treatment. * P < 0.05 and ** P < 0.01 as determined by one-way ANOVA followed by Tukey’s multiple comparison test. e Colony formation post 10 ng/ml EGF treatment in control and Syndecan-1-silenced SUM-149 cells. ** P < 0.01 and # P < 0.001 as determined by one-way ANOVA followed by Tukey’s multiple comparison test. f Western blot analysis of the downstream signaling p-Akt (Ser473) of EGFR signaling in response to EGF stimulation in control and Syndecan-1-silenced SUM-149 cells. Data represent mean ± SEM, n ≥ 3. Data shown are a single experiment representative of three independent experiments

Article Snippet: The human IBC cell line SUM-149 (a kind gift from Dr. Bonnie Sloane, Wayne State University, Detroit, MI, USA) and the non-IBC cell line SKBR3 (ATCC/LGC Promochem, Wesel, Germany) were maintained in HAM’s-F12 and DMEM containing 10% FCS, 1% glutamine and 1% penicillin/streptomycin in a humidified atmosphere of 5% CO 2 at 37 °C, respectively.

Techniques: Expressing, Activation Assay, Transformation Assay, MANN-WHITNEY, Control, Comparison, Western Blot