kat2a primary cell based screening assays cell culture ht1080 human fibrosarcoma cell line ht1080  (ATCC)

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: The Histone Deacetylase Inhibitor Vorinostat Selectively Sensitizes Fibrosarcoma Cells to Chemotherapy

doi: 10.1002/jor.21274

Figure Lengend Snippet: Vorinostat inhibits soft tissue sarcoma and normal fibroblast proliferation. (A) Representative phase-contrast photomicrographs (×200) indicating decreased HT1080, SK-LMS-1, SW872 (STS), and WS-1 normal fibroblast proliferation and altered morphology after vorinostat treatment for 72 h. (B) STS and normal fibroblast proliferation were decreased by vorinostat treatment, as determined by counting cells after 24 h (white bars) or 72 h (black bars). Bars represent the mean and whiskers represent the SEM of at least four biological replicates. # and * denote statistical significance versus cells treated with vehicle after 24 or 72 h, respectively (p < 0.05).

Article Snippet: Cell Culture Human HT1080 fibrosarcoma, SK-LMS-1 leiomyosarcoma, and SW872 liposarcoma cells, and WS-1 normal human fibroblasts were obtained from the ATCC (Manassas, VA) and cultured in minimal essential media (Gibco, Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT), nonessential amino acids (Gibco), sodium pyruvate (Gibco), and penicillin–streptomycin (Gibco) at 37°C under 5% CO 2 .

Techniques:

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: The Histone Deacetylase Inhibitor Vorinostat Selectively Sensitizes Fibrosarcoma Cells to Chemotherapy

doi: 10.1002/jor.21274

Figure Lengend Snippet: Vorinostat does not affect basal apoptosis levels in soft tissue sarcoma cells and normal fibroblasts. STS cells and normal fibroblasts were treated with 1.0 μM vorinostat or vehicle for 24 or 72 h and then trypsinized, fixed, stained for cleaved caspase 3 (BD Pharmingen™) and analyzed by flow cytometry. (A) Representative histograms of cleaved caspase 3 staining in HT1080 fibrosarcoma cells treated with vehicle or vorinostat. The negative control sample (first row, first histogram) was treated with vehicle for 72 h and then stained with the secondary Alexa Fluor® 647 antibody (Invitrogen) only. Puromycin (first row, second histogram) was used to induce apoptosis as a positive control for cleaved caspase 3 staining. (B) Vorinostat only moderately affects caspase 3 cleavage and then only at the supratherapeutic 3.0 μM dose in all cell lines tested. Bars represent the mean and whiskers represent the SEM of at least four biological replicates. # and * denote statistical significance versus cells treated with vehicle after 24 or 72 h, respectively (p < 0.05).

Article Snippet: Cell Culture Human HT1080 fibrosarcoma, SK-LMS-1 leiomyosarcoma, and SW872 liposarcoma cells, and WS-1 normal human fibroblasts were obtained from the ATCC (Manassas, VA) and cultured in minimal essential media (Gibco, Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT), nonessential amino acids (Gibco), sodium pyruvate (Gibco), and penicillin–streptomycin (Gibco) at 37°C under 5% CO 2 .

Techniques: Staining, Flow Cytometry, Negative Control, Positive Control

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: The Histone Deacetylase Inhibitor Vorinostat Selectively Sensitizes Fibrosarcoma Cells to Chemotherapy

doi: 10.1002/jor.21274

Figure Lengend Snippet: Vorinostat sensitizes fibrosarcoma but not normal fibroblasts to doxorubicin. STS cells and normal fibroblasts were treated with 1.0 μM vorinostat or vehicle for 72 h. Doxorubicin was added at the beginning of the third day. (A) Representative (×200) phase-contrast and fluorescent photomicrographs of HT1080 cells plated in chamber slides, treated with vorinostat, doxorubicin, or both and then fixed and stained with anti-cleaved caspase 3 (BD Pharmingen™) and anti-rabbit Alexa Fluor® 488 (Invitrogen) antibodies. I: Phase contrast; II: cleaved caspase 3; III: doxorubicin; IV: DAPI. (B) Vorinostat and doxorubicin synergistically induce apoptosis in HT1080 but not SK-LMS-1 or SW872 STS cells, or WS-1 normal fibroblasts. After a 72-h treatment with vorinostat, doxorubicin, or both, cells were trypsinized, fixed, stained for cleaved caspase 3 and analyzed by flow cytometry. Bars represent the mean and whiskers represent the SEM of at least four biological replicates. Statistical significance is denoted by: * versus vehicle, # versus vorinostat alone, † versus doxorubicin alone.

Article Snippet: Cell Culture Human HT1080 fibrosarcoma, SK-LMS-1 leiomyosarcoma, and SW872 liposarcoma cells, and WS-1 normal human fibroblasts were obtained from the ATCC (Manassas, VA) and cultured in minimal essential media (Gibco, Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT), nonessential amino acids (Gibco), sodium pyruvate (Gibco), and penicillin–streptomycin (Gibco) at 37°C under 5% CO 2 .

Techniques: Staining, Flow Cytometry

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: The Histone Deacetylase Inhibitor Vorinostat Selectively Sensitizes Fibrosarcoma Cells to Chemotherapy

doi: 10.1002/jor.21274

Figure Lengend Snippet: Vorinostat inhibits growth and induces chemosensitivity of fibrosarcoma xenografts. Crl:NU-Foxn1nu nude mice were injected subcutaneously with HT1080 cells. One week after inoculation, mice began receiving daily injections of vorinostat or vehicle and weekly injections of doxorubicin or vehicle. (A,B) HT1080 xenograft growth is inhibited by vorinostat alone and further inhibited by the combination of vorinostat and doxorubicin, but not by doxorubicin alone. Data are represented in (A) as the median and interquartile range and in (B) as the minima and maxima (whiskers) and the median, 25th and 75th percentiles (box) at day 21. N = 16 (Veh, VOR) or N = 9 (Veh + Dox, VOR + Dox). (C) Vorinostat treatment induces histone H3 accumulation in xenografts. Statistical significance is denoted by: *p < 0.05 versus vehicle, #p < 0.01 versus vehicle, †p < 0.01 versus doxorubicin alone.

Article Snippet: Cell Culture Human HT1080 fibrosarcoma, SK-LMS-1 leiomyosarcoma, and SW872 liposarcoma cells, and WS-1 normal human fibroblasts were obtained from the ATCC (Manassas, VA) and cultured in minimal essential media (Gibco, Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT), nonessential amino acids (Gibco), sodium pyruvate (Gibco), and penicillin–streptomycin (Gibco) at 37°C under 5% CO 2 .

Techniques: Injection

Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

Article Title: The Histone Deacetylase Inhibitor Vorinostat Selectively Sensitizes Fibrosarcoma Cells to Chemotherapy

doi: 10.1002/jor.21274

Figure Lengend Snippet: Combined vorinostat and doxorubicin treatment induces cell death in fibrosarcoma xenografts. (A) Cleaved caspase 3 immunohistochemistry revealed that cell death was greatest in HT1080 xenografts from vorinostat/doxorubicin-treated mice (upper panel). The high magnification inset indicates cleaved caspase 3-positive apoptotic cells in the xenograft from vorinostat/doxorubicin-treated mice. TUNEL staining (middle panel) confirmed that vorinostat/doxorubicin induced more apoptosis in HT1080 compared to vehicle, doxorubicin alone, or vorinostat alone. Nuclei were counterstained using DAPI (lower panel). (B) Quantitation of TUNEL staining in HT1080 xenografts. Statistical significance is denoted by *p < 0.05 versus vehicle, #p < 0.05 versus doxorubicin alone, †p < 0.01 versus vorinostat alone. Scale bar = 50 μm.

Article Snippet: Cell Culture Human HT1080 fibrosarcoma, SK-LMS-1 leiomyosarcoma, and SW872 liposarcoma cells, and WS-1 normal human fibroblasts were obtained from the ATCC (Manassas, VA) and cultured in minimal essential media (Gibco, Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT), nonessential amino acids (Gibco), sodium pyruvate (Gibco), and penicillin–streptomycin (Gibco) at 37°C under 5% CO 2 .

Techniques: Immunohistochemistry, TUNEL Assay, Staining, Quantitation Assay

Journal: Biomaterials science

Article Title: X-ray radio-enhancement by Ti 3 C 2 T x MXenes in soft tissue sarcoma.

doi: 10.1039/d3bm00607g

Figure Lengend Snippet: Fig. 4 Surviving fractions of human soft tissue sarcoma cells irradiated at different X-ray doses, relative to untreated, non-irradiated control, treated with (a) fresh Ti3C2Tx MXenes, (b) aged Ti3C2Tx MXenes and (c) TiO2 (P90). (d) Dose-modifying ratio (DMR) at 50% cell survival for afore- mentioned particle types and different concentrations in HT1080 (* indi- cates p < 0.05). (e–g) Surviving fractions of normal human dermal fibro- blasts irradiated at different X-ray doses, relative to untreated, non-irra- diated control, treated with (e) fresh Ti3C2Tx MXenes, (f) aged Ti3C2Tx MXenes and (g) TiO2 (P90).

Article Snippet: T hi s ar tic le is li ce ns ed u nd er a C re at iv e C om m on s A ttr ib ut io nN on C om m er ci al 3 .0 U np or te d L ic en ce Cell lines and culture conditions Human soft tissue sarcoma HT1080 cells (ATCC®CCL121TM) were cultured in minimum essential medium Eagle (MEM, Sigma-Aldrich or Gibco) supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich), 1% L-glutamine (Sigma-Aldrich), 1% non-essential amino acids (NEAA, Sigma-Aldrich), 1% penicillin–streptomycin (PS, Sigma-Aldrich) and 1 mM sodium-pyruvate at 37 °C under a humidified atmosphere containing 5% CO2.

Techniques: Irradiation, Control

Journal: American Journal of Translational Research

Article Title: A novel method for potentiation of chemotherapy in soft tissue sarcomas with BromAc

doi:

Figure Lengend Snippet: 72-hour treatment with doxorubicin. Top graphs represent (A) HT1080, (B) SW872, (C) VAESBJ, and (D) SW982 cell proliferation compared to an untreated control. Significance is denoted by (*) such that P-values are represented by P<0.05 (*), P<0.005 (**), P<0.0005 (***), and P<0.0001 (****). Bottom graphs represent dose-response curve of (E) HT1080, (F) SW872, (G) VAESBJ, and (H) SW982 cell lines with associated IC50, calculated using Prism GraphPad 7.

Article Snippet: Cell culture Human sarcoma cell lines HT1080, SW872, VAESBJ and SW982 were attained from the American Type Culture Collection (ATCC, USA).

Techniques: Control

Journal: American Journal of Translational Research

Article Title: A novel method for potentiation of chemotherapy in soft tissue sarcomas with BromAc

doi:

Figure Lengend Snippet: Efficacy of 72-hour treatment with doxorubicin alone and in combination with bromelain and acetylcysteine in (A) HT1080, (B) SW872, (C) VAESBJ, and (D) SW982 cells measured through proliferation compared to untreated controls. Data are presented as mean ± SD.

Article Snippet: Cell culture Human sarcoma cell lines HT1080, SW872, VAESBJ and SW982 were attained from the American Type Culture Collection (ATCC, USA).

Techniques:

Journal: American Journal of Translational Research

Article Title: A novel method for potentiation of chemotherapy in soft tissue sarcomas with BromAc

doi:

Figure Lengend Snippet: Combination index of doxorubicin in varying combinations with bromelain and acetylcysteine in (A) HT1080, (B) SW872, (C) VAESBJ, and (D) SW982 cell lines. The dotted lines represent thresholds for Chou-Talalay antagonism (CI>1.1) as well as synergy (CI<0.9). Additive effect corresponds to (0.9

Article Snippet: Cell culture Human sarcoma cell lines HT1080, SW872, VAESBJ and SW982 were attained from the American Type Culture Collection (ATCC, USA).

Techniques:

Journal: American Journal of Translational Research

Article Title: A novel method for potentiation of chemotherapy in soft tissue sarcomas with BromAc

doi:

Figure Lengend Snippet: Combination Index of doxorubicin in varying combinations with bromelain and acetylcysteine, displayed for various concentrations of doxorubicin

Article Snippet: Cell culture Human sarcoma cell lines HT1080, SW872, VAESBJ and SW982 were attained from the American Type Culture Collection (ATCC, USA).

Techniques:

Journal: American Journal of Translational Research

Article Title: A novel method for potentiation of chemotherapy in soft tissue sarcomas with BromAc

doi:

Figure Lengend Snippet: Combination index of doxorubicin in varying combinations with bromelain and acetylcysteine, displayed for various concentrations of doxorubicin

Article Snippet: Cell culture Human sarcoma cell lines HT1080, SW872, VAESBJ and SW982 were attained from the American Type Culture Collection (ATCC, USA).

Techniques:

Journal: American Journal of Translational Research

Article Title: A novel method for potentiation of chemotherapy in soft tissue sarcomas with BromAc

doi:

Figure Lengend Snippet: Mucin depletion as assessed quantitatively through ImageJ RGB analysis

Article Snippet: Cell culture Human sarcoma cell lines HT1080, SW872, VAESBJ and SW982 were attained from the American Type Culture Collection (ATCC, USA).

Techniques: Control

Journal: American Journal of Translational Research

Article Title: A novel method for potentiation of chemotherapy in soft tissue sarcomas with BromAc

doi:

Figure Lengend Snippet: Immunofluorescence of soft tissue sarcoma HT1080 cells following 48 hours of treatment with bromelain, acetylcysteine, and bromelain plus acetylcysteine. Expression of MUC1 was viewed under confocal microscope with red and green fluorescence corresponding to nucleus and MUC1 staining, respectively. Scale bar: 50 μm. Final magnification =600×.

Article Snippet: Cell culture Human sarcoma cell lines HT1080, SW872, VAESBJ and SW982 were attained from the American Type Culture Collection (ATCC, USA).

Techniques: Immunofluorescence, Expressing, Microscopy, Fluorescence, Staining

Journal: American Journal of Translational Research

Article Title: A novel method for potentiation of chemotherapy in soft tissue sarcomas with BromAc

doi:

Figure Lengend Snippet: Immunofluorescence of soft tissue sarcoma HT1080 cells following 48 hours of treatment with bromelain, acetylcysteine, and bromelain plus acetylcysteine. Expression of MUC4 was viewed under confocal microscope with red and green fluorescence corresponding to nucleus and MUC4 staining, respectively. Scale bar: 50 μm. Final magnification =600×.

Article Snippet: Cell culture Human sarcoma cell lines HT1080, SW872, VAESBJ and SW982 were attained from the American Type Culture Collection (ATCC, USA).

Techniques: Immunofluorescence, Expressing, Microscopy, Fluorescence, Staining

Journal: American Journal of Translational Research

Article Title: A novel method for potentiation of chemotherapy in soft tissue sarcomas with BromAc

doi:

Figure Lengend Snippet: Western blot analysis of a panel of biomarkers for apoptosis, cell cycle progression, and autophagy in soft tissue sarcoma HT1080 and SW872 cells.

Article Snippet: Cell culture Human sarcoma cell lines HT1080, SW872, VAESBJ and SW982 were attained from the American Type Culture Collection (ATCC, USA).

Techniques: Western Blot

Journal: Molecules

Article Title: Cathepsin B-Cleavable Cyclopeptidic Chemotherapeutic Prodrugs

doi: 10.3390/molecules25184285

Figure Lengend Snippet: Figure 4. HT1080 cell viability after 72 h of incubation with increasing concentrations of cPCP5. Calculated IC50 was 0.86 µM (DOX eq). This result comes from three separated experiments done in triplicates (n = 9). The histogram presented in the top-right corner shows the comparison of the cytotoxicity results in cells incubated for 72 h with increasing concentrations of cPCP4/52, cPCP4/102, cPCP2/20 and cPCP5. (**** p < 0.0001).

Article Snippet: Cell Culture Human fibrosarcoma cell line HT1080 (ATCC® CCL-121TM, Manassas, VA, USA) was grown as monolayers in DMEM (1x) + GlutaMAXTM-1 medium supplemented with 10% fetal calf serum and 100 μL/mL streptomycin and 100 IU/mL penicillin.

Techniques: Incubation, Comparison

Journal: Molecules

Article Title: Cathepsin B-Cleavable Cyclopeptidic Chemotherapeutic Prodrugs

doi: 10.3390/molecules25184285

Figure Lengend Snippet: Figure 5. Confocal microscopy micrographs of HT1080 cells incubated with 30 µM of cPCP4/52 for 24, 48 and 72 h. Control cells are presented after 24 h incubation in the cell medium. DOX fluorescence is presented in red. Hoechst 33342 was used to stain the nuclei (blue) and LysoTracker® Green (LTG) to label the lysosomes (green). Yellow color represents the merged signal between red and green channels meaning the co-localization of DOX with cell lysosomes. Magnification 63X, scale bar 20 µm.

Article Snippet: Cell Culture Human fibrosarcoma cell line HT1080 (ATCC® CCL-121TM, Manassas, VA, USA) was grown as monolayers in DMEM (1x) + GlutaMAXTM-1 medium supplemented with 10% fetal calf serum and 100 μL/mL streptomycin and 100 IU/mL penicillin.

Techniques: Confocal Microscopy, Incubation, Control, Staining

Journal: Journal of cell science

Article Title: Role of membrane-tension gated Ca 2+ flux in cell mechanosensation.

doi: 10.1242/jcs.208470

Figure Lengend Snippet: Fig. 1. Response of HT1080 cells to mechanical compression. (A) Experimental setup for the compression device. Fabrication of the device follows the method in Si et al. (2015). Cells in the device are compressed by the application of air pressure above the PDMS layer. The compression depth is limited by the support pillars with height 4–6 µm. (B) Epifluorescence image of cell (YFP channel) before and after compression, and after the release of compression. The cross-section area of the cell increases under compression, and decreases after the release of compression. Scale bars: 20 µm. (C) Scaled FRET-to-CFP ratio (FRET/CFP) and cell spreading area over time. Compression takes place at t=0 min (indicated by the black arrow). Cells are then released at t=38 min (indicated by the red arrow). Results are mean±s.e.m. (n=40). (D) Summary of mean±s.e.m. FRET/CFP when cell is uncompressed, compressed and released from compression. All results are scaled to the mean value of uncompressed cells and represent three biological replicates (n=40 cells). For each biological replicate, there is one technical replicate. ***P<0.001; n.s., not significant (Student’s t-test).

Article Snippet: Cell culture Human fibrosarcoma HT1080 (ATCC) cells were cultured in high glucose (4.5 g/l) Dulbecco’s modified Eagle’s medium (Mediatech), supplemented with 10% fetal bovine serum (Hyclone) and 1% penicillin-streptomycin (Sigma).

Techniques:

Journal: Journal of cell science

Article Title: Role of membrane-tension gated Ca 2+ flux in cell mechanosensation.

doi: 10.1242/jcs.208470

Figure Lengend Snippet: Fig. 4. Intracellular Ca2+ dye signal after cells are treated with TRPV4 inhibitor or incubated in Ca2+-free medium for 30 min. (A) Image of a cell in the DIC channel and the fluorescence channel showing Ca2+ dye in the cytoplasm. Scale bar: 20 µm. (B) Total intracellular Ca2+ dye signal over time for HT1080 cells with no treatment (control), TRPV4 inhibitor treatment and Ca2+-free medium treatment. The Ca2+ dye signal is generally stable for 15 min after imaging. The slight decrease in signal is due to dye leakage out of the cell, and occurs for all conditions. (C) Population and time average of total intracellular Ca2+ dye signal for control, TRPV4 inhibitor treatment and Ca2+-free medium treatment. Results are mean±s.e.m.; the number of cells in each set of experiments is labeled in C; there were two biological repeats and two technical repeats for each biological repeat in all conditions. ***P<10−9 (Student’s t-test).

Article Snippet: Cell culture Human fibrosarcoma HT1080 (ATCC) cells were cultured in high glucose (4.5 g/l) Dulbecco’s modified Eagle’s medium (Mediatech), supplemented with 10% fetal bovine serum (Hyclone) and 1% penicillin-streptomycin (Sigma).

Techniques: Incubation, Fluorescence, Control, Imaging, Labeling