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: Nature Communications

Article Title: Identification of p62/SQSTM1 as a component of non-canonical Wnt VANGL2–JNK signalling in breast cancer

doi: 10.1038/ncomms10318

Figure Lengend Snippet: ( a ) Basal cell lines chosen for their high basal score/correlation were SUM149 r =0.36 ( a ) and HCC1806 ( b ) r =0.28 and threshold was 0.15. Expression of two short hairpin RNAs abrogated VANGL2 expression in SUM149 cells by western blot analysis (upper panels). SUM149 cells (5 × 10 6 ) were subcutaneously inoculated into the right flank of 4–6-week-old female NSG mice. Tumoral volume was measured at different times (lower panels). The mean and s.e.m. values ( n =6, for shLuc and shVANGL2-transfected cells). The statistical significance between the data sets was determined using a two-way ANOVA test. * P ≤0.05, ** P ≤0.005. ( b ) Same as a using HCC1806 cells, except that 1 × 10 6 cells were inoculated into NSG mice. ( c , d ) Downregulation of VANGL2 with two different shRNAs led to a decreased proliferation of SUM149 ( c ) and HCC1806 ( d ) cells. Error bars represent mean±s.d. ( e ) COMMA-D cells were transduced with lentiviral supernatants allowing expression of GFP or GFP–VANGL2. Cell extracts were probed by western blot analysis with anti-GFP, -VANGL2 and -tubulin antibodies. An asterisk pinpoints endogenous VANGL2. ( f ) Kaplan–Meier curve of tumour-free status of mice transplanted with COMMA-D cells overexpressing GFP or GFP–VANGL2 ( n =30). The statistical significance between the data sets was determined using a log-rank test.

Article Snippet: Second, we tested anchorage-dependent and -independent proliferation and observed that loss of VANGL2 decreased the proliferation rates of SUM149 ( and ) and HCC1806 cells ( ).

Techniques: Expressing, Western Blot, Transfection, Transduction

Journal: Nature Communications

Article Title: Identification of p62/SQSTM1 as a component of non-canonical Wnt VANGL2–JNK signalling in breast cancer

doi: 10.1038/ncomms10318

Figure Lengend Snippet: LC-MS/MS using LTQ-Velos-Orbitrap mass spectrometry analysis of proteins co-immunoprecipitated with VANGL2 in SUM149 cell extracts.

Article Snippet: Second, we tested anchorage-dependent and -independent proliferation and observed that loss of VANGL2 decreased the proliferation rates of SUM149 ( and ) and HCC1806 cells ( ).

Techniques: Mass Spectrometry, Sequencing

Journal: Nature Communications

Article Title: Identification of p62/SQSTM1 as a component of non-canonical Wnt VANGL2–JNK signalling in breast cancer

doi: 10.1038/ncomms10318

Figure Lengend Snippet: ( a ) Endogenous interaction between VANGL2 and p62/SQSTM1 is revealed by western blot analysis after immunoprecipitation using SKBR7 cell extracts. TL is total lysate. Crtl ab is an isotype-matched antibody. ( b ) The VANGL2–p62/SQSTM1 interaction occurs independently of LC3. Proteins were extracted from SUM149 cells and co-immunoprecipitations were carried out with the indicated antibodies. LC3 co-precipitates with p62/SQSTM1 but not with VANGL2. Reciprocally, VANGL2 co-immunoprecipitates with p62/SQSTM1 but not with LC3. IP control antibodies (IP crtl ab) are a polyclonal rabbit (for IP LC3) and a monoclonal rat antibody (for IP VANGL2). IgHs are immunoglobulin heavy chains. ( c ) GST pulldown assays of in vitro translated GFP–VANGL2 (full length: WT, N-terminal 1–102: NT, C-terminal 242–521: CT) showed that VANGL2 WT and CT directly bind to GST–p62/SQSTM1 (GST-p62) but not to GST. Asterisks indicate in vitro translated VANGL2 (top panel) and GST (bottom panel) proteins. AR, autoradiography; CBB, Coomassie Brilliant Blue. ( d ) A p62/SQSTM1 peptide (p62 DN ) disrupts the endogenous VANGL2–p62/SQSTM1 complex. SUM149 cell protein extracts were incubated with the indicated peptides p62 DN or scrambled control peptide (Ctrl peptide) at 100 μM. VANGL2 was then immunoprecipitated (IP VANGL2) and bound proteins were immunoblotted with the indicated antibodies. TLs showed that equal amounts of proteins were present in each condition.

Article Snippet: Second, we tested anchorage-dependent and -independent proliferation and observed that loss of VANGL2 decreased the proliferation rates of SUM149 ( and ) and HCC1806 cells ( ).

Techniques: Western Blot, Immunoprecipitation, Control, In Vitro, Autoradiography, Incubation

Journal: Nature Communications

Article Title: Identification of p62/SQSTM1 as a component of non-canonical Wnt VANGL2–JNK signalling in breast cancer

doi: 10.1038/ncomms10318

Figure Lengend Snippet: ( a ) Downregulation of VANGL2 in SUM149 cells using a specific shRNA led to reduced JNK phosphorylation induced by Wnt5a (100 ng ml −1 for the indicated times). JNK is represented by two isoforms: p54 and p46. Wnt5a led to p46 (phospho-p46) and not p54 (phospho-54) phosphorylation. Relative quantification of immunoblots (phosphorylated JNK/total JNK) is representative from three independent experiments and use of two different shRNAs. ( b ) Expression of GFP-p62 DN , but not GFP, in SUM149 cells led to decreased p46 JNK phosphorylation (phospho-46) induced by 100 ng ml −1 of Wnt5a at the indicated times. Relative quantification of immunoblots (phosphorylated JNK/total JNK) is representative from three independent experiments. ( c ) SUM149 cell extracts were added with the control peptide (Ctrl peptide) or the p62/SQSTM1 peptide (p62 DN ) that inhibited recruitment of JNK and p62/SQSTM1 to VANGL2. ( d ) Proteins extracted from SUM149 cells treated or not with serum were immunoprecipitated with anti-VANGL2 antibody and blotted with the indicated antibodies. p62/SQSTM1, JNK and phosphorylated JNK (phospho-p54 and phospho-p46) were present in the VANGL2 complex.

Article Snippet: Second, we tested anchorage-dependent and -independent proliferation and observed that loss of VANGL2 decreased the proliferation rates of SUM149 ( and ) and HCC1806 cells ( ).

Techniques: shRNA, Phospho-proteomics, Quantitative Proteomics, Western Blot, Expressing, Control, Immunoprecipitation

Journal: International Journal of Medical Sciences

Article Title: Antioxydation And Cell Migration Genes Are Identified as Potential Therapeutic Targets in Basal-Like and BRCA1 Mutated Breast Cancer Cell Lines

doi: 10.7150/ijms.20508

Figure Lengend Snippet: Main characteristics of the cell lines.

Article Snippet: SUM149 and SUM1315 human breast cancer cell lines were obtained from Asterand (Hertfordshire, UK) and grown in Ham's F12 medium according to the manufacturer's instructions.

Techniques: Mutagenesis

Journal: International Journal of Medical Sciences

Article Title: Antioxydation And Cell Migration Genes Are Identified as Potential Therapeutic Targets in Basal-Like and BRCA1 Mutated Breast Cancer Cell Lines

doi: 10.7150/ijms.20508

Figure Lengend Snippet: Summary of transcriptome and methylome data.

Article Snippet: SUM149 and SUM1315 human breast cancer cell lines were obtained from Asterand (Hertfordshire, UK) and grown in Ham's F12 medium according to the manufacturer's instructions.

Techniques: Standard Deviation

Journal: International Journal of Medical Sciences

Article Title: Antioxydation And Cell Migration Genes Are Identified as Potential Therapeutic Targets in Basal-Like and BRCA1 Mutated Breast Cancer Cell Lines

doi: 10.7150/ijms.20508

Figure Lengend Snippet: Comparison of RNA-Seq and immunohistochemistry data.

Article Snippet: SUM149 and SUM1315 human breast cancer cell lines were obtained from Asterand (Hertfordshire, UK) and grown in Ham's F12 medium according to the manufacturer's instructions.

Techniques: Comparison, Immunohistochemistry

Journal: International Journal of Medical Sciences

Article Title: Antioxydation And Cell Migration Genes Are Identified as Potential Therapeutic Targets in Basal-Like and BRCA1 Mutated Breast Cancer Cell Lines

doi: 10.7150/ijms.20508

Figure Lengend Snippet: List of the 77 genes highly expressed and significantly up-regulated in basal-like breast cancer cell lines.

Article Snippet: SUM149 and SUM1315 human breast cancer cell lines were obtained from Asterand (Hertfordshire, UK) and grown in Ham's F12 medium according to the manufacturer's instructions.

Techniques: