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Bristol Myers etoposide vp 16
Induction of H-sema E mRNA in CDDP-sensitive cells. ( a ) Induction of H-sema E in CDDP-sensitive TYKnu cells by CDDP treatment. TYKnu cells were treated for 3 days with 0.0 (untreated control, lane 3), 0.1 (lane 4), 0.3 (lane 5), or 1.0 (lane 6) μg/ml CDDP. The constitutive expression of H-sema E by CDDP-resistant TYKnuR cells (lane 1) was not affected by the 3-day treatment with 1.0 μg/ml CDDP (lane 2). The relative intensity of the blots compared with lane 3 was 5.4 (lane 1), 6.3 (lane 2), 0.2 (lane 4), 1.5 (lane 5), and 4.3 (lane 6). ( b ) Time-dependent induction of H-sema E in CDDP-sensitive Lu65 cells. Lu65 cells were untreated or treated with 2.0 μg/ml CDDP for 6 to 72 hr (hereafter, lanes marked “0” represent untreated controls). The relative intensity of the blots compared with the untreated control was 1.1 (6 hr), 1.0 (24 hr), 2.0 (48 hr), and 3.7 (72 hr). ( c ) Dose-dependent induction of H-sema E in CDDP-sensitive Lu65 cells by the platinum-containing compounds, CBDCA and CDDP. H-sema E was induced in a dose-dependent manner by 3-day treatment with 3–30 μg/ml CBDCA or 0.5–2.0 μg/ml CDDP. The relative intensity of the blots compared with the untreated controls was 1.8 (3 μg/ml CBDCA), 1.8 (10 μg/ml CBDCA), 3.4 (30 μg/ml CBDCA), 1.2 (0.5 μg/ml CDDP), 2.9 (1.0 μg/ml CDDP), and 5.7 (2.0 μg/ml CDDP). ( d ) Induction of H-sema E in CDDP-sensitive Lu65 cells by non-platinum-containing anti-cancer compounds. H-sema E was induced by 3-day treatment with 0.1 and 0.2 μg/ml MMC, 0.1 μg/ml ADM, and 1.0 μg/ml <t>VP-16.</t> The relative intensity of the blots compared with the untreated controls was 2.2 (0.1 μg/ml MMC), 2.7 (0.2 μg/ml MMC), 2.9 (0.1 μg/ml ADM), and 3.1 (1.0 μg/ml VP-16). ( e ) Time- and dose-dependent induction of H-sema E in CDDP-sensitive Lu65 cells by UV irradiation. Total RNA was extracted from Lu65 cells 3 days after UV irradiation at 20, 40, or 80 J/m 2 ( Left ), or 24, 48, or 72 hr after UV irradiation at 80 J/m 2 ( Right ). The relative intensity of the blots compared with the untreated controls was 1.1 (20 J/m 2 ), 1.5 (40 J/m 2 ), 2.5 (80 J/m 2 ), 0.6 (24 hr), 2.0 (48 hr), and 3.2 (72 hr). ( f ) Induction of H-sema E in CDDP-sensitive Lu65 cells by x-ray irradiation. Total RNA was extracted from Lu65 cells 3 days after irradiation at a dose of 3 or 10 Gy. The relative intensity of the blots compared with the untreated control was 0.7 (3 Gy) and 3.9 (10 Gy).
Etoposide Vp 16, supplied by Bristol Myers, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Identification of semaphorin E as a non-MDR drug resistance gene of human cancers"

Article Title: Identification of semaphorin E as a non-MDR drug resistance gene of human cancers

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi:

Induction of H-sema E mRNA in CDDP-sensitive cells. ( a ) Induction of H-sema E in CDDP-sensitive TYKnu cells by CDDP treatment. TYKnu cells were treated for 3 days with 0.0 (untreated control, lane 3), 0.1 (lane 4), 0.3 (lane 5), or 1.0 (lane 6) μg/ml CDDP. The constitutive expression of H-sema E by CDDP-resistant TYKnuR cells (lane 1) was not affected by the 3-day treatment with 1.0 μg/ml CDDP (lane 2). The relative intensity of the blots compared with lane 3 was 5.4 (lane 1), 6.3 (lane 2), 0.2 (lane 4), 1.5 (lane 5), and 4.3 (lane 6). ( b ) Time-dependent induction of H-sema E in CDDP-sensitive Lu65 cells. Lu65 cells were untreated or treated with 2.0 μg/ml CDDP for 6 to 72 hr (hereafter, lanes marked “0” represent untreated controls). The relative intensity of the blots compared with the untreated control was 1.1 (6 hr), 1.0 (24 hr), 2.0 (48 hr), and 3.7 (72 hr). ( c ) Dose-dependent induction of H-sema E in CDDP-sensitive Lu65 cells by the platinum-containing compounds, CBDCA and CDDP. H-sema E was induced in a dose-dependent manner by 3-day treatment with 3–30 μg/ml CBDCA or 0.5–2.0 μg/ml CDDP. The relative intensity of the blots compared with the untreated controls was 1.8 (3 μg/ml CBDCA), 1.8 (10 μg/ml CBDCA), 3.4 (30 μg/ml CBDCA), 1.2 (0.5 μg/ml CDDP), 2.9 (1.0 μg/ml CDDP), and 5.7 (2.0 μg/ml CDDP). ( d ) Induction of H-sema E in CDDP-sensitive Lu65 cells by non-platinum-containing anti-cancer compounds. H-sema E was induced by 3-day treatment with 0.1 and 0.2 μg/ml MMC, 0.1 μg/ml ADM, and 1.0 μg/ml VP-16. The relative intensity of the blots compared with the untreated controls was 2.2 (0.1 μg/ml MMC), 2.7 (0.2 μg/ml MMC), 2.9 (0.1 μg/ml ADM), and 3.1 (1.0 μg/ml VP-16). ( e ) Time- and dose-dependent induction of H-sema E in CDDP-sensitive Lu65 cells by UV irradiation. Total RNA was extracted from Lu65 cells 3 days after UV irradiation at 20, 40, or 80 J/m 2 ( Left ), or 24, 48, or 72 hr after UV irradiation at 80 J/m 2 ( Right ). The relative intensity of the blots compared with the untreated controls was 1.1 (20 J/m 2 ), 1.5 (40 J/m 2 ), 2.5 (80 J/m 2 ), 0.6 (24 hr), 2.0 (48 hr), and 3.2 (72 hr). ( f ) Induction of H-sema E in CDDP-sensitive Lu65 cells by x-ray irradiation. Total RNA was extracted from Lu65 cells 3 days after irradiation at a dose of 3 or 10 Gy. The relative intensity of the blots compared with the untreated control was 0.7 (3 Gy) and 3.9 (10 Gy).
Figure Legend Snippet: Induction of H-sema E mRNA in CDDP-sensitive cells. ( a ) Induction of H-sema E in CDDP-sensitive TYKnu cells by CDDP treatment. TYKnu cells were treated for 3 days with 0.0 (untreated control, lane 3), 0.1 (lane 4), 0.3 (lane 5), or 1.0 (lane 6) μg/ml CDDP. The constitutive expression of H-sema E by CDDP-resistant TYKnuR cells (lane 1) was not affected by the 3-day treatment with 1.0 μg/ml CDDP (lane 2). The relative intensity of the blots compared with lane 3 was 5.4 (lane 1), 6.3 (lane 2), 0.2 (lane 4), 1.5 (lane 5), and 4.3 (lane 6). ( b ) Time-dependent induction of H-sema E in CDDP-sensitive Lu65 cells. Lu65 cells were untreated or treated with 2.0 μg/ml CDDP for 6 to 72 hr (hereafter, lanes marked “0” represent untreated controls). The relative intensity of the blots compared with the untreated control was 1.1 (6 hr), 1.0 (24 hr), 2.0 (48 hr), and 3.7 (72 hr). ( c ) Dose-dependent induction of H-sema E in CDDP-sensitive Lu65 cells by the platinum-containing compounds, CBDCA and CDDP. H-sema E was induced in a dose-dependent manner by 3-day treatment with 3–30 μg/ml CBDCA or 0.5–2.0 μg/ml CDDP. The relative intensity of the blots compared with the untreated controls was 1.8 (3 μg/ml CBDCA), 1.8 (10 μg/ml CBDCA), 3.4 (30 μg/ml CBDCA), 1.2 (0.5 μg/ml CDDP), 2.9 (1.0 μg/ml CDDP), and 5.7 (2.0 μg/ml CDDP). ( d ) Induction of H-sema E in CDDP-sensitive Lu65 cells by non-platinum-containing anti-cancer compounds. H-sema E was induced by 3-day treatment with 0.1 and 0.2 μg/ml MMC, 0.1 μg/ml ADM, and 1.0 μg/ml VP-16. The relative intensity of the blots compared with the untreated controls was 2.2 (0.1 μg/ml MMC), 2.7 (0.2 μg/ml MMC), 2.9 (0.1 μg/ml ADM), and 3.1 (1.0 μg/ml VP-16). ( e ) Time- and dose-dependent induction of H-sema E in CDDP-sensitive Lu65 cells by UV irradiation. Total RNA was extracted from Lu65 cells 3 days after UV irradiation at 20, 40, or 80 J/m 2 ( Left ), or 24, 48, or 72 hr after UV irradiation at 80 J/m 2 ( Right ). The relative intensity of the blots compared with the untreated controls was 1.1 (20 J/m 2 ), 1.5 (40 J/m 2 ), 2.5 (80 J/m 2 ), 0.6 (24 hr), 2.0 (48 hr), and 3.2 (72 hr). ( f ) Induction of H-sema E in CDDP-sensitive Lu65 cells by x-ray irradiation. Total RNA was extracted from Lu65 cells 3 days after irradiation at a dose of 3 or 10 Gy. The relative intensity of the blots compared with the untreated control was 0.7 (3 Gy) and 3.9 (10 Gy).

Techniques Used: Expressing, Irradiation

Related Articles

Modification:

Article Title: Increased sensitivity to gemcitabine of P-glycoprotein and multidrug resistance-associated protein-overexpressing human cancer cell lines
Article Snippet: .. Chemicals and reagents Dulbecco's modified eagle's medium (DMEM) and Rosswell Park Memorial Institute (RPMI) medium were purchased from Flow Laboratories (Irvine, UK); foetal calf serum from Life Technologies (New York, NY, USA); trichloroacetic acid (TCA), glutamine and gentamicin from Merck (Darmstadt, Germany); trypsin and sulphorhodamine B (SRB) from Sigma Chemical Co. (St Louis, USA); and VP-16 (etoposide) from Bristol-Myers Squibb (Weesp, the Netherlands). .. Eli Lilly (Indianapolis, IN, USA) kindly supplied gemcitabine and [5-3 H]gemcitabine (16.7 Ci mmol−1 ).

Concentration Assay:

Article Title: Melphalan exposure induces an Interleukin-6 deficit in bone marrow stromal cells and osteoblasts
Article Snippet: .. Etoposide (VP-16) (Bristol-Myers Squibb, New York, NY) was stored at a concentration of 33.98 mM and a final concentration of 50 μM was used in all experiments. .. Methotrexate (Sigma; [50μg/ml]), vincristine (Sigma; [20μg/ml]), docetaxel (Sigma; [50μM]), carboplatin (Sigma; [50μM]) or mechlorethamine hydrochloride (Sigma; [10μM or 25μM]) were included where indicated.

other:

Article Title: Modulation of Nuclear Factor E2-related Factor-2 (Nrf2) Activation by the Stress Response Gene Immediate Early Response-3 (IER3) in Colonic Epithelial Cells
Article Snippet: was from Calbiochem, tBHQ and SFN from Sigma, Killer-TRAIL from Enzo Life-Science/Alexis (Lörrach, Germany), and etoposide (Vepesid) from Bristol-Myers/Squibb.

Sulforhodamine B Assay:

Article Title: Increased sensitivity to gemcitabine of P-glycoprotein and multidrug resistance-associated protein-overexpressing human cancer cell lines
Article Snippet: .. Chemicals and reagents Dulbecco's modified eagle's medium (DMEM) and Rosswell Park Memorial Institute (RPMI) medium were purchased from Flow Laboratories (Irvine, UK); foetal calf serum from Life Technologies (New York, NY, USA); trichloroacetic acid (TCA), glutamine and gentamicin from Merck (Darmstadt, Germany); trypsin and sulphorhodamine B (SRB) from Sigma Chemical Co. (St Louis, USA); and VP-16 (etoposide) from Bristol-Myers Squibb (Weesp, the Netherlands). .. Eli Lilly (Indianapolis, IN, USA) kindly supplied gemcitabine and [5-3 H]gemcitabine (16.7 Ci mmol−1 ).

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    Bristol Myers vepesid
    <t>Etoposide</t> and ANS-etoposide toxicity in GSTA1 and MGST1 overexpressing cells. Cellular toxicity of (A) etoposide and (B) ANS–etoposide analyzed with the MTT assay after short-term 24 h exposure with indicated doses, in at least three independent experiments measured in at least triplicate (mean ± SEM, n ≥ 9, * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001). Left panel: MGST1 overexpressing MCF7 cells (dark gray/MGST1) and the vector control (light gray/control). Right panel: GSTA1 overexpressing V79 cells (dark gray/GSTA1) and the vector control (light gray/control).
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    Etoposide and ANS-etoposide toxicity in GSTA1 and MGST1 overexpressing cells. Cellular toxicity of (A) etoposide and (B) ANS–etoposide analyzed with the MTT assay after short-term 24 h exposure with indicated doses, in at least three independent experiments measured in at least triplicate (mean ± SEM, n ≥ 9, * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001). Left panel: MGST1 overexpressing MCF7 cells (dark gray/MGST1) and the vector control (light gray/control). Right panel: GSTA1 overexpressing V79 cells (dark gray/GSTA1) and the vector control (light gray/control).

    Journal: Molecular pharmaceutics

    Article Title: Chemical Reactivity Window Determines Prodrug Efficiency toward Glutathione Transferase Overexpressing Cancer Cells

    doi: 10.1021/acs.molpharmaceut.6b00140

    Figure Lengend Snippet: Etoposide and ANS-etoposide toxicity in GSTA1 and MGST1 overexpressing cells. Cellular toxicity of (A) etoposide and (B) ANS–etoposide analyzed with the MTT assay after short-term 24 h exposure with indicated doses, in at least three independent experiments measured in at least triplicate (mean ± SEM, n ≥ 9, * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001). Left panel: MGST1 overexpressing MCF7 cells (dark gray/MGST1) and the vector control (light gray/control). Right panel: GSTA1 overexpressing V79 cells (dark gray/GSTA1) and the vector control (light gray/control).

    Article Snippet: 1-Chloro-2,4-dinitrobenzene (CDNB), reduced glutathione, and doxorubicin were purchased from Sigma-Aldrich, whereas etoposide was purchased from Vepesid, Bristol-Myers.

    Techniques: MTT Assay, Plasmid Preparation

    Chemical structures and conversion of prodrugs. (A) Chemical structures of DOX and etoposide as well as the prodrugs MNS–DOX, DNS-DOX, ANS–OX, and ANS–etoposide are depicted. (B) Conversion of the prodrug yields a free doxorubicin/etoposide, sulfur dioxide and an acetylmononitro/dinitro benzene moiety conjugated to GSH. These prodrugs are hypothesized to enter the cell via a passive diffusion. In the cell, GSTs are able to activate the prodrugs via their sulfonamidase and sulfonamide cleavage activity.

    Journal: Molecular pharmaceutics

    Article Title: Chemical Reactivity Window Determines Prodrug Efficiency toward Glutathione Transferase Overexpressing Cancer Cells

    doi: 10.1021/acs.molpharmaceut.6b00140

    Figure Lengend Snippet: Chemical structures and conversion of prodrugs. (A) Chemical structures of DOX and etoposide as well as the prodrugs MNS–DOX, DNS-DOX, ANS–OX, and ANS–etoposide are depicted. (B) Conversion of the prodrug yields a free doxorubicin/etoposide, sulfur dioxide and an acetylmononitro/dinitro benzene moiety conjugated to GSH. These prodrugs are hypothesized to enter the cell via a passive diffusion. In the cell, GSTs are able to activate the prodrugs via their sulfonamidase and sulfonamide cleavage activity.

    Article Snippet: 1-Chloro-2,4-dinitrobenzene (CDNB), reduced glutathione, and doxorubicin were purchased from Sigma-Aldrich, whereas etoposide was purchased from Vepesid, Bristol-Myers.

    Techniques: Diffusion-based Assay, Activity Assay

    Synthesis of ANS–DOX (1) and ANS–Etoposide (2); See Materials and Methods for Details

    Journal: Molecular pharmaceutics

    Article Title: Chemical Reactivity Window Determines Prodrug Efficiency toward Glutathione Transferase Overexpressing Cancer Cells

    doi: 10.1021/acs.molpharmaceut.6b00140

    Figure Lengend Snippet: Synthesis of ANS–DOX (1) and ANS–Etoposide (2); See Materials and Methods for Details

    Article Snippet: 1-Chloro-2,4-dinitrobenzene (CDNB), reduced glutathione, and doxorubicin were purchased from Sigma-Aldrich, whereas etoposide was purchased from Vepesid, Bristol-Myers.

    Techniques:

    Relative quantification of DOX-responder microRNAs in rat heart across all groups. Relative quantification of (A) miR-208b, (B) miR-215, (C) miR-216b, (D) miR-367 and (E) miR-34c in DOX, DOX + DZR, EPS groups, normalized versus vehicle treated animals. Expression levels were measured by single assay qPCR (n = 3, except #, n = 2). DOX: Doxorubicin, DZR: dexrazoxane, EPS: etoposide; numbers indicate the weekly dose of each compound in mg/kg/week. Empty spaces represent non-sampled animals. The vehicle treated is the first column of each time-point. The animals used in this experiment were distinct from the ones represented in Table 1 . Error bars represent SD. T-test results are indicated by asterisks for significant DOX-treated groups vs. their own vehicle-treated, unless otherwise specified by horizontal range bars; *P

    Journal: PLoS ONE

    Article Title: Perturbation of microRNAs in Rat Heart during Chronic Doxorubicin Treatment

    doi: 10.1371/journal.pone.0040395

    Figure Lengend Snippet: Relative quantification of DOX-responder microRNAs in rat heart across all groups. Relative quantification of (A) miR-208b, (B) miR-215, (C) miR-216b, (D) miR-367 and (E) miR-34c in DOX, DOX + DZR, EPS groups, normalized versus vehicle treated animals. Expression levels were measured by single assay qPCR (n = 3, except #, n = 2). DOX: Doxorubicin, DZR: dexrazoxane, EPS: etoposide; numbers indicate the weekly dose of each compound in mg/kg/week. Empty spaces represent non-sampled animals. The vehicle treated is the first column of each time-point. The animals used in this experiment were distinct from the ones represented in Table 1 . Error bars represent SD. T-test results are indicated by asterisks for significant DOX-treated groups vs. their own vehicle-treated, unless otherwise specified by horizontal range bars; *P

    Article Snippet: Doxorubicin (Adriamycin, Lot No. 86G23FY, CAS# 23214-92-8), a generous gift from Adria Laboratories Inc. (Columbus, OH, USA), etoposide (ETOPOPHOS® Lot No. 5E04155, CAS# 117091-64-2) was purchased from Bristol-Myers Squibb (Princeton, NJ, USA) and dexrazoxane (Zinecard® Lot No. ADR074B, CAS# 24584-09-6) purchased from Pharmacia/Pfizer (Kalamazoo, MI, USA) were administered once weekly as follows: 11 week old male Crl:CD(SD) rats received DOX or EPS via intravenous injection once a week for 2, 4, or 6 weeks based on at a dose volume of 2 mL/kg.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Study design and representative micrograph showing DOX-related vacuolation in the myocardium. (A) Six adult male rats were injected with the indicated doses of vehicle, doxorubicin (DOX), dexrazoxane (DZR), etoposide (EPS) or a combination of DOX and DZR for 2, 4 or 6 weeks. Cardiac tissue was excised and deep frozen for gene expression and microRNA profiling experiments. A representative micrograph of a toluidine blue stained myocardial section of a control (B) and of a DOX treated animal (C). Black arrows indicate sarcoplasmic micro- and macro- vacuolation of cardiomyocytes.

    Journal: PLoS ONE

    Article Title: Perturbation of microRNAs in Rat Heart during Chronic Doxorubicin Treatment

    doi: 10.1371/journal.pone.0040395

    Figure Lengend Snippet: Study design and representative micrograph showing DOX-related vacuolation in the myocardium. (A) Six adult male rats were injected with the indicated doses of vehicle, doxorubicin (DOX), dexrazoxane (DZR), etoposide (EPS) or a combination of DOX and DZR for 2, 4 or 6 weeks. Cardiac tissue was excised and deep frozen for gene expression and microRNA profiling experiments. A representative micrograph of a toluidine blue stained myocardial section of a control (B) and of a DOX treated animal (C). Black arrows indicate sarcoplasmic micro- and macro- vacuolation of cardiomyocytes.

    Article Snippet: Doxorubicin (Adriamycin, Lot No. 86G23FY, CAS# 23214-92-8), a generous gift from Adria Laboratories Inc. (Columbus, OH, USA), etoposide (ETOPOPHOS® Lot No. 5E04155, CAS# 117091-64-2) was purchased from Bristol-Myers Squibb (Princeton, NJ, USA) and dexrazoxane (Zinecard® Lot No. ADR074B, CAS# 24584-09-6) purchased from Pharmacia/Pfizer (Kalamazoo, MI, USA) were administered once weekly as follows: 11 week old male Crl:CD(SD) rats received DOX or EPS via intravenous injection once a week for 2, 4, or 6 weeks based on at a dose volume of 2 mL/kg.

    Techniques: Injection, Expressing, Staining

    Structure of podophyllotoxin derivatives used in this study. Structure of F14512. Epipodophyllopoxin is shown to the left, with the substitution of different R groups leading to teniposide (VM26), TOP-53 and etoposide (VP16).

    Journal: PLoS ONE

    Article Title: An Integrated Drosophila Model System Reveals Unique Properties for F14512, a Novel Polyamine-Containing Anticancer Drug That Targets Topoisomerase II

    doi: 10.1371/journal.pone.0023597

    Figure Lengend Snippet: Structure of podophyllotoxin derivatives used in this study. Structure of F14512. Epipodophyllopoxin is shown to the left, with the substitution of different R groups leading to teniposide (VM26), TOP-53 and etoposide (VP16).

    Article Snippet: Drugs Etoposide, TOP-53 and F14512 were obtained from Pierre Fabre Medicament and teniposide from Bristol-Myers Squibb.

    Techniques:

    Suppression of white-mottled PEV by Topo II poisons. Photographs show representative eyes of 5-day old w m4h females hatched from late L3 larvae fed etoposide (VP16), F14512 or teniposide (VM26) at the indicated concentrations. A comparison of the no-drug controls shown in the left column highlights the intrinsic heterogeneity of the white-mottled phenotype. Etoposide and F14512 partly suppress white PEV at higher concentrations and display an apparent threshold effect at 25 µM or 50 µM, respectively. Teniposide feeding leads to full dose-dependent suppression of PEV. Controls ( w m4h or wild-type Oregon R larvae fed 0 or 1% DMSO used to dilute the drugs, a concentration that corresponds to that used in 100-µM drug treatments) are shown in the bottom row. The graph at the bottom shows the corresponding quantifications of extracted eye pigment and represents mean values obtained from three independent experiments performed in duplicate (30 5-day old females scored for each condition). Results are represented as % pigment relative to Oregon R wild-type (WT) flies fed 1% DMSO. Error bars represent the experimental error. Note that the toxicity of F14512 feeding at 25 and 50 µM only permitted analysis of 10 females for each experiment, most likely accounting for the substantial variations observed (p

    Journal: PLoS ONE

    Article Title: An Integrated Drosophila Model System Reveals Unique Properties for F14512, a Novel Polyamine-Containing Anticancer Drug That Targets Topoisomerase II

    doi: 10.1371/journal.pone.0023597

    Figure Lengend Snippet: Suppression of white-mottled PEV by Topo II poisons. Photographs show representative eyes of 5-day old w m4h females hatched from late L3 larvae fed etoposide (VP16), F14512 or teniposide (VM26) at the indicated concentrations. A comparison of the no-drug controls shown in the left column highlights the intrinsic heterogeneity of the white-mottled phenotype. Etoposide and F14512 partly suppress white PEV at higher concentrations and display an apparent threshold effect at 25 µM or 50 µM, respectively. Teniposide feeding leads to full dose-dependent suppression of PEV. Controls ( w m4h or wild-type Oregon R larvae fed 0 or 1% DMSO used to dilute the drugs, a concentration that corresponds to that used in 100-µM drug treatments) are shown in the bottom row. The graph at the bottom shows the corresponding quantifications of extracted eye pigment and represents mean values obtained from three independent experiments performed in duplicate (30 5-day old females scored for each condition). Results are represented as % pigment relative to Oregon R wild-type (WT) flies fed 1% DMSO. Error bars represent the experimental error. Note that the toxicity of F14512 feeding at 25 and 50 µM only permitted analysis of 10 females for each experiment, most likely accounting for the substantial variations observed (p

    Article Snippet: Drugs Etoposide, TOP-53 and F14512 were obtained from Pierre Fabre Medicament and teniposide from Bristol-Myers Squibb.

    Techniques: Concentration Assay

    Evidence for F14512-specific Topo II-mediated cleavage in moderately repeated Drosophila sequences. (A) Hin dIII-digested DNA samples from S2 cells treated for 30 minutes with no drug (lanes 1and 7), F14512 (5, 10, 25 or 50 µM, lanes 2–6 and 9–12) or 50 µM TOP-53 (lanes 2 and 8), in the absence (lanes 1–7) or presence (lanes 8–12) of 25 µM distamycin (DIST). Purified DNA samples were hybridized to a labeled Hin fI-digested whole genomic DNA probe that had been pre-hybridized as described in Materials. The a–b–c fragments indicated to the right of the gel are described in the text. (B) Densitometric scan of the autoradiograph shown in (A) for lanes 7, 8 and 11. Regions corresponding to fragments a, b and c are indicated by arrowheads. (C) S2 cells were treated for 30 minutes with no drug (-), F14512 (25 or 50 µM) or 50 µM etoposide in the presence of 25 µM distamycin (DIST). Purified DNA samples were processed as described in panel A. Fragments a, b and c are indicated by arrowheads to the left of the gel, which provides better resolution of the a–b regions.

    Journal: PLoS ONE

    Article Title: An Integrated Drosophila Model System Reveals Unique Properties for F14512, a Novel Polyamine-Containing Anticancer Drug That Targets Topoisomerase II

    doi: 10.1371/journal.pone.0023597

    Figure Lengend Snippet: Evidence for F14512-specific Topo II-mediated cleavage in moderately repeated Drosophila sequences. (A) Hin dIII-digested DNA samples from S2 cells treated for 30 minutes with no drug (lanes 1and 7), F14512 (5, 10, 25 or 50 µM, lanes 2–6 and 9–12) or 50 µM TOP-53 (lanes 2 and 8), in the absence (lanes 1–7) or presence (lanes 8–12) of 25 µM distamycin (DIST). Purified DNA samples were hybridized to a labeled Hin fI-digested whole genomic DNA probe that had been pre-hybridized as described in Materials. The a–b–c fragments indicated to the right of the gel are described in the text. (B) Densitometric scan of the autoradiograph shown in (A) for lanes 7, 8 and 11. Regions corresponding to fragments a, b and c are indicated by arrowheads. (C) S2 cells were treated for 30 minutes with no drug (-), F14512 (25 or 50 µM) or 50 µM etoposide in the presence of 25 µM distamycin (DIST). Purified DNA samples were processed as described in panel A. Fragments a, b and c are indicated by arrowheads to the left of the gel, which provides better resolution of the a–b regions.

    Article Snippet: Drugs Etoposide, TOP-53 and F14512 were obtained from Pierre Fabre Medicament and teniposide from Bristol-Myers Squibb.

    Techniques: Purification, Labeling, Autoradiography

    Growth inhibition of Drosophila Kc cells by F14512 and characteristic cytotoxic effects. (A) Antiproliferative properties of F14512 and reference Topo II poisons, measured through viable cell counting (Vi-CELL) in exponentially growing Drosophila Kc cells. Drugs were used at the following concentrations: F14512 1 µM (-□-) and 50 µM (-▪-); etoposide (VP16) 1 µM (-Δ-) and 50 µM (-▴-); and TOP-53 1 µM (-○-) and 50 µM (-•-); control cells (-◊-) were treated with 0.1% DMSO alone. Viability was measured after the treatment times shown. Mean values +SD of three independent experiments are reported. (B) Phase-contrast micrographs of Kc cells treated as shown with etoposide (VP16), TOP-53 and F14512 and photographed after 24 hours. Arrowheads point to aggregates of apoptotic debris (etoposide VP16, TOP-53) or to elongated and/or multinucleated cells (F14512). Bar: 5 µm. Note that F14512 treatment yields no evident sign of apoptosis at the concentrations used here. (C) Phase-contrast or fluorescence micrographs of intact Kc cells and DAPI-stained fixed cells, respectively, after 24 hours of treatment with etoposide (VP16) or F14512 at the concentrations shown. The black arrowhead in the etoposide-treated photograph indicates apoptotic bodies, DAPI-stained samples show signs of hypercondensed chromatin. Low concentrations of F14512 (1 or 5 µM) again yield large adherent, elongated and multinucleated cells, DAPI staining shows evidence of internuclear chromatin bridges (white arrowheads) Bar: 5 µm.

    Journal: PLoS ONE

    Article Title: An Integrated Drosophila Model System Reveals Unique Properties for F14512, a Novel Polyamine-Containing Anticancer Drug That Targets Topoisomerase II

    doi: 10.1371/journal.pone.0023597

    Figure Lengend Snippet: Growth inhibition of Drosophila Kc cells by F14512 and characteristic cytotoxic effects. (A) Antiproliferative properties of F14512 and reference Topo II poisons, measured through viable cell counting (Vi-CELL) in exponentially growing Drosophila Kc cells. Drugs were used at the following concentrations: F14512 1 µM (-□-) and 50 µM (-▪-); etoposide (VP16) 1 µM (-Δ-) and 50 µM (-▴-); and TOP-53 1 µM (-○-) and 50 µM (-•-); control cells (-◊-) were treated with 0.1% DMSO alone. Viability was measured after the treatment times shown. Mean values +SD of three independent experiments are reported. (B) Phase-contrast micrographs of Kc cells treated as shown with etoposide (VP16), TOP-53 and F14512 and photographed after 24 hours. Arrowheads point to aggregates of apoptotic debris (etoposide VP16, TOP-53) or to elongated and/or multinucleated cells (F14512). Bar: 5 µm. Note that F14512 treatment yields no evident sign of apoptosis at the concentrations used here. (C) Phase-contrast or fluorescence micrographs of intact Kc cells and DAPI-stained fixed cells, respectively, after 24 hours of treatment with etoposide (VP16) or F14512 at the concentrations shown. The black arrowhead in the etoposide-treated photograph indicates apoptotic bodies, DAPI-stained samples show signs of hypercondensed chromatin. Low concentrations of F14512 (1 or 5 µM) again yield large adherent, elongated and multinucleated cells, DAPI staining shows evidence of internuclear chromatin bridges (white arrowheads) Bar: 5 µm.

    Article Snippet: Drugs Etoposide, TOP-53 and F14512 were obtained from Pierre Fabre Medicament and teniposide from Bristol-Myers Squibb.

    Techniques: Inhibition, Cell Counting, Fluorescence, Staining

    Cellular profiles of drug-induced Topo II-mediated cleavage in SAT III repeats of the Drosophila X chromosome. (A) Structure of SAT III repeats. 359-bp SAT III repeats accommodate 2 nucleosomes per repeat. One of the two nucleosomal linkers located in each repeat contains a short GC-rich sequence that serves as a highly specific target for Topo II cleavage in vivo , as denoted by scissors. (B) Schneider S2 cells were treated for 30 minutes with teniposide (VM26), etoposide (VP16) or TOP-53 at the concentrations shown, in the absence (-) or presence (+) of distamycin (DIST, 25 µM). Purified DNA was electrophoresed without prior restriction enzyme digestion, transferred to a nylon membrane and hybridized to a labeled SAT III probe. 359-bp: the SAT III monomer (SAT) used as a probe; M: molecular weight standards. ( C ) S2 cells were treated for 30 minutes with no drug (0, lane 13), F14512 (5, 10, 25 or 50 µM, lanes 14–17) or 50 µM TOP-53 (lane 18), in the presence of 25 µM distamycin (DIST). Omission of the latter gave identical results (not shown). DNA samples were processed as described above. SAT: the 359-bp cloned fragment used as a probe; M: molecular weight standards.

    Journal: PLoS ONE

    Article Title: An Integrated Drosophila Model System Reveals Unique Properties for F14512, a Novel Polyamine-Containing Anticancer Drug That Targets Topoisomerase II

    doi: 10.1371/journal.pone.0023597

    Figure Lengend Snippet: Cellular profiles of drug-induced Topo II-mediated cleavage in SAT III repeats of the Drosophila X chromosome. (A) Structure of SAT III repeats. 359-bp SAT III repeats accommodate 2 nucleosomes per repeat. One of the two nucleosomal linkers located in each repeat contains a short GC-rich sequence that serves as a highly specific target for Topo II cleavage in vivo , as denoted by scissors. (B) Schneider S2 cells were treated for 30 minutes with teniposide (VM26), etoposide (VP16) or TOP-53 at the concentrations shown, in the absence (-) or presence (+) of distamycin (DIST, 25 µM). Purified DNA was electrophoresed without prior restriction enzyme digestion, transferred to a nylon membrane and hybridized to a labeled SAT III probe. 359-bp: the SAT III monomer (SAT) used as a probe; M: molecular weight standards. ( C ) S2 cells were treated for 30 minutes with no drug (0, lane 13), F14512 (5, 10, 25 or 50 µM, lanes 14–17) or 50 µM TOP-53 (lane 18), in the presence of 25 µM distamycin (DIST). Omission of the latter gave identical results (not shown). DNA samples were processed as described above. SAT: the 359-bp cloned fragment used as a probe; M: molecular weight standards.

    Article Snippet: Drugs Etoposide, TOP-53 and F14512 were obtained from Pierre Fabre Medicament and teniposide from Bristol-Myers Squibb.

    Techniques: Sequencing, In Vivo, Purification, Labeling, Molecular Weight, Clone Assay

    Chemotherapeutic drug-induced cell cycle arrest with down-regulation of Akt kinase activity. (A) 293T or HeLa cells were treated with medium alone (cont.) or 10 μM VP-16, 5 μM cDDP, or 100 nM 17-AAG. After treatment for 24 h, cellular DNA content was determined by a flow cytometer. (B) After treatment with the indicated chemotherapeutic drugs as for panel A for 4 h, cell lysates were subjected to immunoblot analysis with the indicated antibodies. (C and D) After treatment with the indicated chemotherapeutic drugs as for panel A for 6 h, endogenous Akt or Cdc2 was immunoprecipitated from cell lysates and was subjected to the Akt or Cdc2 kinase assay as described in Materials and Methods. Each vertical bar represents the mean ± standard deviation of three independent experiments. (E) 293T cells were transfected with nonsilencing control siRNA (cont.) or WEE1Hu siRNAs (WEE1Hu-1 and WEE1Hu-2). After transfection for 48 h, cells were treated with the indicated chemotherapeutic drugs as for panel A for 4 h. The cell lysates were subjected to immunoblot analysis with the indicated antibodies. (F and G) 293T cells were transfected with nonsilencing control siRNA (cont.) or WEE1Hu siRNA (WEE1Hu-1). After transfection for 48 h, cells were treated with the indicated chemotherapeutic drugs as for panel A for 6 h. Endogenous Akt or Cdc2 was immunoprecipitated from cell lysates and was subjected to the Akt or Cdc2 kinase assay as described in Materials and Methods. Each vertical bar represents the mean ± standard deviation of three independent experiments.

    Journal: Molecular and Cellular Biology

    Article Title: Akt/Protein Kinase B-Dependent Phosphorylation and Inactivation of WEE1Hu Promote Cell Cycle Progression at G2/M Transition

    doi: 10.1128/MCB.25.13.5725-5737.2005

    Figure Lengend Snippet: Chemotherapeutic drug-induced cell cycle arrest with down-regulation of Akt kinase activity. (A) 293T or HeLa cells were treated with medium alone (cont.) or 10 μM VP-16, 5 μM cDDP, or 100 nM 17-AAG. After treatment for 24 h, cellular DNA content was determined by a flow cytometer. (B) After treatment with the indicated chemotherapeutic drugs as for panel A for 4 h, cell lysates were subjected to immunoblot analysis with the indicated antibodies. (C and D) After treatment with the indicated chemotherapeutic drugs as for panel A for 6 h, endogenous Akt or Cdc2 was immunoprecipitated from cell lysates and was subjected to the Akt or Cdc2 kinase assay as described in Materials and Methods. Each vertical bar represents the mean ± standard deviation of three independent experiments. (E) 293T cells were transfected with nonsilencing control siRNA (cont.) or WEE1Hu siRNAs (WEE1Hu-1 and WEE1Hu-2). After transfection for 48 h, cells were treated with the indicated chemotherapeutic drugs as for panel A for 4 h. The cell lysates were subjected to immunoblot analysis with the indicated antibodies. (F and G) 293T cells were transfected with nonsilencing control siRNA (cont.) or WEE1Hu siRNA (WEE1Hu-1). After transfection for 48 h, cells were treated with the indicated chemotherapeutic drugs as for panel A for 6 h. Endogenous Akt or Cdc2 was immunoprecipitated from cell lysates and was subjected to the Akt or Cdc2 kinase assay as described in Materials and Methods. Each vertical bar represents the mean ± standard deviation of three independent experiments.

    Article Snippet: Etoposide (VP-16) and cisplatin (cDDP) were kindly provided by Bristol-Myers Squibb Co., Ltd. (Tokyo, Japan). were purchased from Sigma (St. Louis, MO).

    Techniques: Activity Assay, Flow Cytometry, Cytometry, Immunoprecipitation, Kinase Assay, Standard Deviation, Transfection