Journal: Veterinary Sciences

Article Title: Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

doi: 10.3390/vetsci8090194

Figure Lengend Snippet: Tumor growth parameters of IPC-366 and SUM149 cell lines in ectopic and orthotopic models.

Article Snippet: The human triple-negative inflammatory breast cancer cell line SUM149 was obtained from Asterand, Inc. (Detroit, MI, USA), (RRID: CVCL_3422).

Techniques: Injection

Journal: Veterinary Sciences

Article Title: Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

doi: 10.3390/vetsci8090194

Figure Lengend Snippet: IPC-366 and SUM149 xenotransplanted mice, paraffin sections, H-E. ( A ) IPC-366 ectopic xenotransplanted mice. Neoplastic cells arranged in solid masses separated by a scant fibrovascular stroma infiltrating the adjacent dermis (inset: neoplastic cells infiltrating adjacent dermis). ( B ) IPC-366 orthotopic mice. Unencapsulated and densely cellular mass extending into the adjacent adipose tissue. ( C , D ) Ectopic and orthotopic IPC-366 xenotransplanted mice. Tumors are composed of highly pleomorphic cells with marked anisocytosis and anisokaryosis. Binucleated cells are commonly seen (arrow). ( E , F ) Ectopic and orthotopic SUM149 xenografted mice. Solid tumors infiltrate the dermis and adipose tissue. No histological differences were found between the types of SUM149 xenografts. ( G ) Orthotopic SUM149 xenograft. Medium to large round cells with a moderate eosinophilic cytoplasm and large nuclei with one or more evident nucleoli. ( H ) Orthotopic SUM149 xenograft. Presence of neoplastic cells with an elongated and empty cytoplasm that displaced the nuclei to the periphery, suggestive of endothelial-like cells (ELCs) (arrow). Atypical mitoses were frequently seen (arrowhead).

Article Snippet: The human triple-negative inflammatory breast cancer cell line SUM149 was obtained from Asterand, Inc. (Detroit, MI, USA), (RRID: CVCL_3422).

Techniques:

Journal: Veterinary Sciences

Article Title: Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

doi: 10.3390/vetsci8090194

Figure Lengend Snippet: Estrogen receptor (ER), Progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER-2) expression on ectopic and orthotopic xenografts from IPC-366 and SUM149 cell lines.

Article Snippet: The human triple-negative inflammatory breast cancer cell line SUM149 was obtained from Asterand, Inc. (Detroit, MI, USA), (RRID: CVCL_3422).

Techniques: Expressing

Journal: Veterinary Sciences

Article Title: Tumor Growth Progression in Ectopic and Orthotopic Xenografts from Inflammatory Breast Cancer Cell Lines

doi: 10.3390/vetsci8090194

Figure Lengend Snippet: Steroid hormone secretion studied (P4, DHEA, A4, T, DHT, E1SO4, and E2), on ectopic (subcutaneous) and orthotopic (mammary fat pad) models of IPC-366 and SUM149 xenografts.

Article Snippet: The human triple-negative inflammatory breast cancer cell line SUM149 was obtained from Asterand, Inc. (Detroit, MI, USA), (RRID: CVCL_3422).

Techniques:

Journal: Cell metabolism

Article Title: Targeting Breast Cancer Stem Cell State Equilibrium through Modulation of Redox Signaling

doi: 10.1016/j.cmet.2018.06.006

Figure Lengend Snippet: (A–E) E- and M-BCSCs in SUM149 (A), HCC1806 (B), MCF7 (C) and T47D (D) BC cells treated with 2DG (10 and 20 mM, 40h) and effects of 2DG to induce apoptosis in M-BCSCs or bulk tumor cells (E). *, **: P<0.05 or 0.01 (vs. no 2DG).

Article Snippet: Cell Labeling and Flow Cytometry To determine the effect of drug treatment on M- and E-BCSCs in basal BC cell lines, SUM149 or HCC1806 following various treatment were digested by 0.25% Trypsin-EDTA, re-suspended in HF buffer (HBSS plus 2% FBS) at 2×10 5 cells/100 μl and incubated with antibodies against human CD24 (PE-Cy7-conjugated, 1:100 from BD for SUM149 or 1:75 from Biolegend for HCC1806) and CD44 (APC-conjugated, 1:200, from BD) in cold room for 30 minutes.

Techniques:

Journal: Cell metabolism

Article Title: Targeting Breast Cancer Stem Cell State Equilibrium through Modulation of Redox Signaling

doi: 10.1016/j.cmet.2018.06.006

Figure Lengend Snippet: (A, B) SUM149 BC cells treated with 2DG/H2O2 for 1.5h (A) or NAC for 20h (B) was analyzed by p-ACC (S79) and HIF1α antibodies.

Article Snippet: Cell Labeling and Flow Cytometry To determine the effect of drug treatment on M- and E-BCSCs in basal BC cell lines, SUM149 or HCC1806 following various treatment were digested by 0.25% Trypsin-EDTA, re-suspended in HF buffer (HBSS plus 2% FBS) at 2×10 5 cells/100 μl and incubated with antibodies against human CD24 (PE-Cy7-conjugated, 1:100 from BD for SUM149 or 1:75 from Biolegend for HCC1806) and CD44 (APC-conjugated, 1:200, from BD) in cold room for 30 minutes.

Techniques:

Journal: Cell metabolism

Article Title: Targeting Breast Cancer Stem Cell State Equilibrium through Modulation of Redox Signaling

doi: 10.1016/j.cmet.2018.06.006

Figure Lengend Snippet: (A–C) Contents of E- (A) and M- (B) BCSCs in SUM149 treated with AUR, BSO or AUR+BSO with or without NAC for 24h and the effects of Trig to sensitize E-BCSCs to AUR (C). *, **: P<0.05 or 0.01 vs. untreated. NS: not significant.

Article Snippet: Cell Labeling and Flow Cytometry To determine the effect of drug treatment on M- and E-BCSCs in basal BC cell lines, SUM149 or HCC1806 following various treatment were digested by 0.25% Trypsin-EDTA, re-suspended in HF buffer (HBSS plus 2% FBS) at 2×10 5 cells/100 μl and incubated with antibodies against human CD24 (PE-Cy7-conjugated, 1:100 from BD for SUM149 or 1:75 from Biolegend for HCC1806) and CD44 (APC-conjugated, 1:200, from BD) in cold room for 30 minutes.

Techniques:

Journal: Cell metabolism

Article Title: Targeting Breast Cancer Stem Cell State Equilibrium through Modulation of Redox Signaling

doi: 10.1016/j.cmet.2018.06.006

Figure Lengend Snippet: (A, B) SUM149 treated with AUR (0.5 μM) or BSO (1mM) for 24h were labeled with Annexin V and examined for the content of E-BCSCs (A) and Annexin V+ cell ratio in ALDH+ and ALDH− cell populations (B).

Article Snippet: Cell Labeling and Flow Cytometry To determine the effect of drug treatment on M- and E-BCSCs in basal BC cell lines, SUM149 or HCC1806 following various treatment were digested by 0.25% Trypsin-EDTA, re-suspended in HF buffer (HBSS plus 2% FBS) at 2×10 5 cells/100 μl and incubated with antibodies against human CD24 (PE-Cy7-conjugated, 1:100 from BD for SUM149 or 1:75 from Biolegend for HCC1806) and CD44 (APC-conjugated, 1:200, from BD) in cold room for 30 minutes.

Techniques: Labeling

Journal: Cell metabolism

Article Title: Targeting Breast Cancer Stem Cell State Equilibrium through Modulation of Redox Signaling

doi: 10.1016/j.cmet.2018.06.006

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cell Labeling and Flow Cytometry To determine the effect of drug treatment on M- and E-BCSCs in basal BC cell lines, SUM149 or HCC1806 following various treatment were digested by 0.25% Trypsin-EDTA, re-suspended in HF buffer (HBSS plus 2% FBS) at 2×10 5 cells/100 μl and incubated with antibodies against human CD24 (PE-Cy7-conjugated, 1:100 from BD for SUM149 or 1:75 from Biolegend for HCC1806) and CD44 (APC-conjugated, 1:200, from BD) in cold room for 30 minutes.

Techniques: Recombinant, Inhibition, Luciferase, Reporter Gene Assay, RNA Sequencing Assay, shRNA, Software

Journal: Oncotarget

Article Title: EGFR signaling promotes inflammation and cancer stem-like activity in inflammatory breast cancer

doi: 10.18632/oncotarget.18958

Figure Lengend Snippet: ( A ) EGFR expression is depleted in SUM149 cells after EGFR knockdown. Expression of EGFR in the stable shCtrl clone and EGFR knockdown clones, shEGFR-1 and shEGFR-3, was analyzed by Western blotting. ( B ) EGFR depletion reduces the formation of mammospheres in SUM149 cells. Bars, ± SD. * P < 0.001. ( C and D ). EGFR depletion decreases the CD44 + /CD24 −/low population (C) and ALDH activity (D) of SUM149 cells. * P < 0.001; ** P < 0.01. ( E ) Erlotinib treatment decreases mammosphere formation of SUM149 cells. Shown are representative images of primary mammospheres. * P < 0.001; ** P = 0.001. ( F ) and ( G ) Erlotinib treatment decreases the CD44 + /CD24 −/low population (F) and ALDH activity (G) of SUM149 cells. FITC, fluorescein isothiocyanate; PE, phycoerythrin. * P < 0.01. ( H ) Panitumumab (PmAb) treatment inhibits EGF-stimulated phosphorylation of EGFR in SUM149 cells. Cells were pretreated with PmAb at the indicated doses for 1 hour and then stimulated with EGF (50 ng/mL) for 15 minutes. ( I ) PmAb treatment decreases the CD44 + /CD24 −/low population of SUM149 cells. ( J ) PmAb treatment decreases the mammosphere formation of SUM149 cells. * P < 0.001; ** P < 0.005. Experiments were independently repeated 3 times.

Article Snippet: The human IBC cell lines SUM149 and SUM190 were purchased from Asterand and were grown in Ham's F12 medium supplemented with 5% FBS, 5 μg/mL insulin, 1 μg/mL hydrocortisone, and 1% antibiotic-antimycotic.

Techniques: Expressing, Knockdown, Clone Assay, Western Blot, Activity Assay, Phospho-proteomics

Journal: Oncotarget

Article Title: EGFR signaling promotes inflammation and cancer stem-like activity in inflammatory breast cancer

doi: 10.18632/oncotarget.18958

Figure Lengend Snippet: ( A ) EGF stimulation increases the expression of COX-2 in SUM149 cells. Cells were serum-starved for 24 hours prior to stimulation with recombinant EGF at 100 ng/mL for the indicated time periods. Ctrl, control. ( B ) EGFR knockdown reduces the expression of COX-2 in SUM149 cells. ( C ) Erlotinib or PmAb treatment reduces the expression of COX-2 in SUM149 cells. ( D ) EGFR and COX-2 mRNA expression correlates in 25 IBC patient biopsy samples. ( E ) In a different population of 44 IBC patients, high COX-2 expression in tumor specimens correlates with worse overall survival. P = 0.0184. ( F ) IBC cells have higher levels of COX-2's enzymatic products, PGE 2 and PGF 2α , than non-IBC cells. Experiments in panels A, B, and C were independently repeated 3 times.

Article Snippet: The human IBC cell lines SUM149 and SUM190 were purchased from Asterand and were grown in Ham's F12 medium supplemented with 5% FBS, 5 μg/mL insulin, 1 μg/mL hydrocortisone, and 1% antibiotic-antimycotic.

Techniques: Expressing, Recombinant, Control, Knockdown

Journal: Oncotarget

Article Title: EGFR signaling promotes inflammation and cancer stem-like activity in inflammatory breast cancer

doi: 10.18632/oncotarget.18958

Figure Lengend Snippet: ( A ) and ( B ) PGE 2 and PGF 2α treatment increases the CD44 + /CD24 −/low population (A) and ALDH activity (B) of SUM149 cells. Cells were treated with 0.5 μM PGE 2 or PGF 2α for 48 hours and then subjected to flow cytometry analysis. * P = 0.01; ** P = 0.001. ( C ) Treatment with COX-2 inhibitor celecoxib decreases ALDH activity of SUM149 cells. Cells were treated with celecoxib at the indicated doses for 48 hours and then subjected to flow cytometry analysis. * P = 0.001; ** P < 0.001. ( D ) Celecoxib treatment decreases mammosphere formation of SUM149 cells. * P < 0.005. ( E ) A stabilized PGE 2 analogue, dmPGE 2 (100 nM), mitigates the decrease of primary (left) and secondary (right) mammosphere formation in SUM149 EGFR-knockdown clone shEGFR-1. * P < 0.005; ** P = 0.01. Experiments were independently repeated 3 times.

Article Snippet: The human IBC cell lines SUM149 and SUM190 were purchased from Asterand and were grown in Ham's F12 medium supplemented with 5% FBS, 5 μg/mL insulin, 1 μg/mL hydrocortisone, and 1% antibiotic-antimycotic.

Techniques: Activity Assay, Flow Cytometry, Knockdown

Journal: Oncotarget

Article Title: EGFR signaling promotes inflammation and cancer stem-like activity in inflammatory breast cancer

doi: 10.18632/oncotarget.18958

Figure Lengend Snippet: ( A ) Celecoxib treatment reduces the mesenchymal phenotype in SUM149 cells. ( B ) Celecoxib treatment reduces projection formation of SUM149 cells. Cells were plated in Matrigel culture with or without celecoxib for 48 hours. Left panel: Representative images of SUM149 cells. Right panel: Projections quantitated by S.CORE analysis. ( C ) PGE 2 (0.5 μM) and PGF 2α (0.5 μM) treatment increases the migration (left panel) and invasion (right panel) of SUM149 cells. * P < 0.001; ** P < 0.005. ( D ) Celecoxib treatment at the indicated dose for 48 hours decreases the migration (left panel) and invasion (right panel) of SUM149 cells. * P < 0.01. ( E ) Celecoxib treatment inhibits tumor growth in a SUM149 xenograft model. Each data point represents the mean tumor volume of eight mice per group. Cxb, celecoxib. Bars, SD. * P = 0.02; ** P = 0.001. ( F ) Celecoxib treatment affects the expression of COX-2, E-cadherin, and vimentin in tumor tissues of SUM149 xenografts. Scale bar, 100 μm. ( G ) Celecoxib treatment reduces the expression of PGE 2 and PGF 2α in SUM149 xenografts. * P = 0.01. ( H ) Celecoxib treatment affects the mesenchymal phenotype in SUM149 xenografts. Experiments in panels A, B, C, D, G, and H were independently repeated 3 times.

Article Snippet: The human IBC cell lines SUM149 and SUM190 were purchased from Asterand and were grown in Ham's F12 medium supplemented with 5% FBS, 5 μg/mL insulin, 1 μg/mL hydrocortisone, and 1% antibiotic-antimycotic.

Techniques: Migration, Expressing

Journal: Oncotarget

Article Title: EGFR signaling promotes inflammation and cancer stem-like activity in inflammatory breast cancer

doi: 10.18632/oncotarget.18958

Figure Lengend Snippet: ( A ) and ( B ) The COX-2 pathway regulates Nodal mRNA in SUM149 cells. SUM149 cells were treated with 0.5 μM PGE 2 or PGF 2α (A) or celecoxib at the indicated doses (B) for 48 hours under 3D culture conditions, and the expression level of Nodal was measured by real-time RT-PCR. ( C ) PGE 2 stimulation activates the Nodal pathway in SUM149 cells. Serum-starved SUM149 cells were stimulated with 0.5 μM PGE 2 for different time periods, and the expression of the indicated proteins was analyzed with Western blotting. ( D ) Celecoxib (10 μM) treatment reduces Nodal expression and inhibits the Nodal pathway in SUM149 cells. ( E ) and ( F ) Recombinant Nodal (rNodal) mitigates the celecoxib-induced inhibition of migration (E) and invasion (F) of SUM149 cells. Cells were treated with different combinations of celecoxib (25 μM) and rNodal (100 ng/mL) as indicated for 48 hours and then a transwell migration or Matrigel invasion assay was performed. * P < 0.001; ** P < 0.05. ( G ) rNodal mitigates the celecoxib-induced inhibition of mammosphere formation of SUM149 cells. * P < 0.01; ** P = 0.001. Experiments were independently repeated 3 times.

Article Snippet: The human IBC cell lines SUM149 and SUM190 were purchased from Asterand and were grown in Ham's F12 medium supplemented with 5% FBS, 5 μg/mL insulin, 1 μg/mL hydrocortisone, and 1% antibiotic-antimycotic.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Recombinant, Inhibition, Migration, Invasion Assay

Journal: Oncotarget

Article Title: EGFR signaling promotes inflammation and cancer stem-like activity in inflammatory breast cancer

doi: 10.18632/oncotarget.18958

Figure Lengend Snippet: ( A ) EGF stimulation activates the Nodal pathway in SUM149 cells. Serum-starved SUM149 cells were stimulated with 50 ng/mL EGF for different time periods, and the expression of the indicated proteins was analyzed with Western blotting. ( B ) Erlotinib (2.5 μM) treatment reduces Nodal expression and inhibits the Nodal pathway in SUM149 cells. ( C ) PmAb (20 μg/mL) treatment reduces Nodal expression and inhibits the Nodal pathway in SUM149 cells. Experiments were independently repeated 3 times.

Article Snippet: The human IBC cell lines SUM149 and SUM190 were purchased from Asterand and were grown in Ham's F12 medium supplemented with 5% FBS, 5 μg/mL insulin, 1 μg/mL hydrocortisone, and 1% antibiotic-antimycotic.

Techniques: Expressing, Western Blot