Journal: mAbs

Article Title: Structural and functional characterization of MBS301, an afucosylated bispecific anti-HER2 antibody

doi: 10.1080/19420862.2018.1486946

Figure Lengend Snippet: ADCC activity evaluation of anti-HER2 antibodies on four target cancer cells.

Article Snippet: Human breast cancer cell line BT-474 (HTB-20), MDA-MB-175VII (HTB-25), SK-BR-3(HTB-30) and HCC1419(CRL-2326) are HER2-positive cell lines, which were obtained from the ATCC, and HCC1419 is trastuzumab-resistant human breast cancer cell line.

Techniques: Activity Assay

Journal: Cancers

Article Title: In Vitro Comparative Study of Near-Infrared Photoimmunotherapy and Photodynamic Therapy

doi: 10.3390/cancers15133400

Figure Lengend Snippet: Cellular binding/uptake specificity and distribution of mAb-IR700 and talaporfin sodium. Fluorescence images of live cells stained for nuclei (blue) after incubation with mAb-IR700 or talaporfin sodium (magenta). ( A ) A431 cells, ( B ) BALB/3T3 cells, and ( C ) BT-474 cells. Scale bar = 20 µm.

Article Snippet: The human breast ductal carcinoma cell line BT-474 (HTB-20) was obtained from the American Type Culture Collection through Summit Pharmaceuticals International Corporation (Tokyo, Japan).

Techniques: Binding Assay, Fluorescence, Staining, Incubation

Journal: Cancers

Article Title: In Vitro Comparative Study of Near-Infrared Photoimmunotherapy and Photodynamic Therapy

doi: 10.3390/cancers15133400

Figure Lengend Snippet: Evaluation of cell death induced by NIR-PIT and TS-PDT. Hoechst and PI double-stained images 1 day after irradiation. ( A ) A431 cells, ( B ) BALB/3T3 cells, and ( C ) BT-474 cells. Scale bar = 50 µm. ( D ) LDH assay at 1 day after irradiation in the three cell lines. Data are presented as means ± SD (n = 3; Tukey’s test; * p < 0.05).

Article Snippet: The human breast ductal carcinoma cell line BT-474 (HTB-20) was obtained from the American Type Culture Collection through Summit Pharmaceuticals International Corporation (Tokyo, Japan).

Techniques: Staining, Irradiation, Lactate Dehydrogenase Assay

Journal: Cancers

Article Title: In Vitro Comparative Study of Near-Infrared Photoimmunotherapy and Photodynamic Therapy

doi: 10.3390/cancers15133400

Figure Lengend Snippet: Analysis of ICD markers following NIR-PIT or TS-PDT. ( A ) Confocal microscopy images of HMGB1 and quantification of HMGB1 translocation. HMGB1 was stained at 1 h after treatment of A431 and BT-474 cells with NIR-PIT or TS-PDT. Scale bar = 20 µm. Fluorescence intensity of nuclear HMGB1 was quantified; data are presented as means ± SD from five microscopic fields (Tukey’s test; * p < 0.05). ( B ) ATP assays of culture supernatants. Data are presented as means ± SD (n = 3; Steel–Dwass test).

Article Snippet: The human breast ductal carcinoma cell line BT-474 (HTB-20) was obtained from the American Type Culture Collection through Summit Pharmaceuticals International Corporation (Tokyo, Japan).

Techniques: Confocal Microscopy, Translocation Assay, Staining, Fluorescence

Journal: Molecular Cancer

Article Title: Phosphorylation of Ser 78 of Hsp27 correlated with HER-2/ neu status and lymph node positivity in breast cancer

doi: 10.1186/1476-4598-6-52

Figure Lengend Snippet: Effect of heregulin α1 (HRG α1) and p38 MAPK inhibitor (SB 203580) on Hsp27 phosphorylation . Cultures of cells from the BT474 breast cancer cell line were treated with HRG α1 for 10 and 30 min (A) or SB 203580 for 10 hours (B) and total cell lysates were extracted using M-PER reagent (Pierce). Equal amounts of proteins (20 μg) were separated by SDS-PAGE and transferred onto PVDF membrane. The membranes were blocked for 1 hour, followed by being probed with the respective primary antibodies (anti-Hsp27, anti-pSer 15 , anti-pSer 78 and anti-pSer 82 ), and HRP-conjugated secondary antibody. The signals were captured and their intensities were detected as described in Figure 2. The phosphorylation levels of pSer 15 , pSer 78 and pSer 82 were expressed as the ratios of intensity probed with phosphorylation site-specific antibody to the intensity probed with anti-Hsp27. Data with ± SD represents the average of triplicate experiments. C: control; SB: inhibitor SB203580. For the control of HRG-treated cells, untreated cells were cultured for 10 and 30 min and equal amounts of cellular proteins from both time intervals were mixed and used as control. For the control of inhibitor-treated cells, cells were treated with DMSO for 10 hours and cellular proteins were used as the control. * p < 0.05 (student t -test)

Article Snippet: The human BT474 breast cancer cell line was obtained from American Type Culture Collection, and maintained in modified Dulbecco's medium (HybriCare) supplemented with 10% fetal bovine serum (FBS) at 37°C in a humidified atmosphere of 5% CO 2 in an incubator.

Techniques: SDS Page, Cell Culture

Journal: BMC Cancer

Article Title: CD44 isoforms are heterogeneously expressed in breast cancer and correlate with tumor subtypes and cancer stem cell markers

doi: 10.1186/1471-2407-11-418

Figure Lengend Snippet: Comparison of the proportion of CD44+ cells as determined by flow cytometry to the relative mRNA expression determined by q-RT-PCR.

Article Snippet: The breast cell lines BT-474, HCC1428, HCC1937, MCF7, MCF10A, MDA-MB-231, MDA-MB-361, MDA-MB-436, SK-BR-3 and ZR-75-1 were obtained from American Type Culture Collection (ATCC, Mannanas, VA).

Techniques: Flow Cytometry, Expressing

Journal: Breast Cancer Research : BCR

Article Title: β1 integrin mediates an alternative survival pathway in breast cancer cells resistant to lapatinib

doi: 10.1186/bcr2936

Figure Lengend Snippet: Phosphorylated levels of β1 downstream kinases are increased upon acquisition of resistance to lapatinib (L) . ( A ) Parental (P) BT474 and ( B ) HCC1954 cells resistant to lapatinib (LRes), trastuzumab (TRes), and combination (LTRes) treatment strategies were developed by long-term exposure in 2D. Protein extracts were probed for β1, pHER2, pFAK, and pSrc, as well as totals.

Article Snippet: The human breast cancer cell line BT474 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained as previously described [ ].

Techniques:

Journal: Breast Cancer Research : BCR

Article Title: β1 integrin mediates an alternative survival pathway in breast cancer cells resistant to lapatinib

doi: 10.1186/bcr2936

Figure Lengend Snippet: β1 blockade overcomes lapatinib resistance in BT474 cells in 3D culture . ( A ) Parental and LRes cells were plated on lrECM and treated ± lapatinib. ( B ) The β1 inhibitory antibody AIIB2--or IgG control--was applied to parental and LRes cells on Day 0 of plating on lrECM and allowed to grow for 12 days. ( C ) Parental and LRes cells were plated in lrECM with appropriate inhibitors, allowed to grow for five days, stained with Ki67 antigen or TUNEL labeling, and imaged. ( D ) 3D cultures of BT474 parental and LRes cells treated with AIIB2 or IgG were allowed to propagate for five days, then harvested for protein and immunoblotting. Densitometry measurements were normalized to total levels--except β1, which was normalized to β-actin--and are representative of three independent experiments. ( E ) The FAK inhibitor PF 573228 was applied to parental and LRes cells on Day 0 of plating on lrECM and cultures were allowed to grow for 12 days.

Article Snippet: The human breast cancer cell line BT474 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained as previously described [ ].

Techniques: Staining, TUNEL Assay, Labeling, Western Blot

Journal: Breast Cancer Research : BCR

Article Title: β1 integrin mediates an alternative survival pathway in breast cancer cells resistant to lapatinib

doi: 10.1186/bcr2936

Figure Lengend Snippet: Percent growth inhibition of cells in response to HER-targeted therapies

Article Snippet: The human breast cancer cell line BT474 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained as previously described [ ].

Techniques: Inhibition

Journal: Breast Cancer Research : BCR

Article Title: β1 integrin mediates an alternative survival pathway in breast cancer cells resistant to lapatinib

doi: 10.1186/bcr2936

Figure Lengend Snippet: HER2 and the β1 pathway play alternate roles in resistance to lapatinib-containing regimens, in comparison to trastuzumab . ( A ) Parental, LRes, and TRes cells were plated on lrECM in the presence of lapatinib and/or trastuzumab and assayed for response. ( B ) 3D cultures of parental, LRes, and TRes BT474 cells were harvested for protein and probed for phosphorylated and total HER receptors.

Article Snippet: The human breast cancer cell line BT474 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained as previously described [ ].

Techniques:

Journal: Breast Cancer Research : BCR

Article Title: β1 integrin mediates an alternative survival pathway in breast cancer cells resistant to lapatinib

doi: 10.1186/bcr2936

Figure Lengend Snippet: β1 inhibition impedes colony grow of BT474 LRes and LTRes cells, but not parental or TRes . ( A ) Parental, LRes, LTRes, and TRes cells were plated in lrECM, subjected to HER2 and/or β1 inhibitors on Day 0, and propagated for 10 to 12 days. ( B ) The Hs_ITGB1_5 siRNA was validated both in 2D and 3D (top), transfected at 40 nMsi into parental, LRes, LTRes, and TRes cells, which were then grown on lrECM for 10 days (bottom).

Article Snippet: The human breast cancer cell line BT474 was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained as previously described [ ].

Techniques: Inhibition, Transfection

Journal: Oncoscience

Article Title: FS-93, an Hsp90 inhibitor, induces G2/M arrest and apoptosis via the degradation of client proteins in oncogene addicted and derived resistant cancer cells

doi:

Figure Lengend Snippet: A. Effects of FS-93 on cell proliferation. BT-474, NCI-H3122 and EBC-1 cells were treated with FS-93 at 25, 50 and 100nM or NVP-AUY922 at 100 nM for 72 h. Cell viability was assessed with SRB assay. Bars represent means±SD. B. Effects of FS-93 on degradation of client onco-proteins. BT-474, NCI-H3122 and EBC-1 cells were treated with FS-93 at 25, 50 and 100nM or NVP-AUY922 at 100 nM for 24 h. Cell lysates were prepared and analyzed with immunoblotting.

Article Snippet: Human breast cancer cell line BT-474 was obtained from American Type Culture Collection (Manassas, VA).

Techniques: Sulforhodamine B Assay, Western Blot

Journal: Oncoscience

Article Title: FS-93, an Hsp90 inhibitor, induces G2/M arrest and apoptosis via the degradation of client proteins in oncogene addicted and derived resistant cancer cells

doi:

Figure Lengend Snippet: A ., B. Effects of FS-93 on cell cycle distribution. BT-474, NCI-H3122 and EBC-1 cells were treated with FS-93 at 25, 50 and 100nM or NVP-AUY922 at 100 nM for 24 h. Cell cycle distribution was analyzed by FACS after propidium iodide staining. Representative images A. and quantification results B. were presented. Bars represent means±SD. C. Impacts of FS-93 on G2/M transition regulators. BT-474, NCI-H3122 and EBC-1 cells were treated with FS-93 at 25, 50 and 100nM or NVP-AUY922 at 100 nM for 24 h and cells lysates were immunoblotted with the indicated antibodies.

Article Snippet: Human breast cancer cell line BT-474 was obtained from American Type Culture Collection (Manassas, VA).

Techniques: Staining

Journal: Oncoscience

Article Title: FS-93, an Hsp90 inhibitor, induces G2/M arrest and apoptosis via the degradation of client proteins in oncogene addicted and derived resistant cancer cells

doi:

Figure Lengend Snippet: A. Effects of FS-93 on apoptosis induction. BT-474, NCI-H3122 and EBC-1 cells were treated with DMSO or FS-93 at 100 nM for 12, 24, 48 and 72h. Apoptosis was determined with annexin-V/PI assay. Bars represent means±SD. B. Impacts of FS-93 on apoptotic proteins. BT-474 cells were treated with DMSO or FS-93 at 100 nM for 12, 24, 48 and 72h and analyzed by immunoblotted with the indicated antibodies.

Article Snippet: Human breast cancer cell line BT-474 was obtained from American Type Culture Collection (Manassas, VA).

Techniques:

Journal: bioRxiv

Article Title: Native size exclusion chromatography-based mass spectrometry (SEC-MS) identifies novel components of the Heat Shock Protein 90-dependent proteome

doi: 10.1101/2022.05.23.492985

Figure Lengend Snippet: (A) Left, Box/Tukey plots illustrate that total levels of the three IDH3 protein complex subunits do not change following HSP90 inhibition. Right, SEC profiles identify a high molecular weight peak (corresponding to the size of the octameric IDH3 complex) that is significantly reduced for all three subunits in HSP90 inhibitor-treated cells. Asterisks depict significantly differential fractions. ( B ) IDH3 activity is significantly reduced upon HSP90 inhibition in HT29 human colon adenocarcinoma, HCT116 human colon carcinoma, and BT474 human breast ductal carcinoma cell lines. Activity was measured using the IDH Activity Assay Kit (Sigma) according to manufacturer’s instructions, using NAD+ as the co-factor. See for the corresponding assay with NADP+ as the co-factor, for estimating IDH1 and IDH2 activity. Adjusted p -values (two-tailed Student’s t -test) are shown.

Article Snippet: HT29 human colon adenocarcinoma, HCT116 human colon carcinoma, and BT474 human breast ductal carcinoma cells purchased from ATCC (LGC Promochem) were cultured in DMEM (Invitrogen) and supplemented with 10 % fetal calf serum (PAA Lab-oratories), 2 mm L-glutamine, 0.1 mm non-essential amino acids and 100 U of penicillin and streptomycin (all from Invitrogen) at 37 °C in a humidified incubator with 5 % CO 2 and sub-cultured at 70 % confluency.

Techniques: Inhibition, Molecular Weight, Activity Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Native size exclusion chromatography-based mass spectrometry (SEC-MS) identifies novel components of the Heat Shock Protein 90-dependent proteome

doi: 10.1101/2022.05.23.492985

Figure Lengend Snippet: ( A ) Combined activities of IDH1 & IDH2 are not significantly reduced upon HSP90 inhibition in HT29 colon adenocarcinoma, HCT116 colon carcinoma, and BT474 breast ductal carcinoma cell lines. Activity was measured using the IDH Activity Assay Kit (Sigma) according to manufacturer’s instructions, using NADP + as the co-factor—which is used by both IDH1 and IDH2. See for corresponding assay with NAD + as the co-factor, for estimating IDH3 activity. ( B ) IDH3 activity is significantly reduced upon treatment with the chemotypically distinct HSP90 inhibitors luminespib (AUY922) and BIIB021, in HCT116 colon carcinoma cells. Activity was measured using the IDH Activity Assay Kit (Sigma) according to manufacturer’s instructions, using NAD + as the co-factor.

Article Snippet: HT29 human colon adenocarcinoma, HCT116 human colon carcinoma, and BT474 human breast ductal carcinoma cells purchased from ATCC (LGC Promochem) were cultured in DMEM (Invitrogen) and supplemented with 10 % fetal calf serum (PAA Lab-oratories), 2 mm L-glutamine, 0.1 mm non-essential amino acids and 100 U of penicillin and streptomycin (all from Invitrogen) at 37 °C in a humidified incubator with 5 % CO 2 and sub-cultured at 70 % confluency.

Techniques: Inhibition, Activity Assay