HOSEpiC (Catalog No.7310) was obtained from ScienCell Research Laboratories (San Diego, CA). OAW42 cells were purchased from the Cell Banks in Cancer Institute and Cancer Hospital, Chinese Academy of Medical Sciences (Beijing, China). The human ovarian adenocarcinoma cell lines, OVCAR3, CAOV3, and SKOV3, were provided by the China Center for Type Culture Collection (Wuhan, China). OvCa cell lines (A2780 and JHOS4) and cisplatin (DDP)-resistant OvCa lines (SKOV3/DDP and CAOV3/DDP) were purchased from the Cell Biology of the Chinese Academy of Sciences (Shanghai, China). DDP, Adriamycin (ADM), and Paclitaxel (PTX) were purchased from Sigma Corporation. Bafilomycin A1 (ab120497) and Recombinant VEGFA (ab117230) were purchased from Abcam Corporation. 3- (4, 5- dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide [MTT, (M2128)] was obtained from the Sigma Corporation and used as a measure of cell viability. B27 (17,504–044), epidermal growth factor (RP-8661), and bFGF (RP-8628) were purchased from Invitrogen. Heparin (9041–08-1) was purchased from Sigma-Aldrich.
Adriamycin adm
Manufactured by Merck Group
Sourced in United States
Adriamycin (ADM) is a chemotherapeutic agent used in the treatment of various types of cancer. It is a red-colored, crystalline powder that is soluble in water and alcohol. Adriamycin is a member of the anthracycline class of antineoplastic drugs and functions by intercalating with DNA, inhibiting DNA and RNA synthesis, and generating free radicals that damage cellular components.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
5 fluorouracil 5 fu, by Merck Group (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Heparin, by Merck Group (1 mentions)
Paclitaxel ptx, by Merck Group (1 mentions)
Ovcar3, by National Collection of Authenticated Cell Cultures (1 mentions)
Caov3, by National Collection of Authenticated Cell Cultures (1 mentions)
Skov3, by National Collection of Authenticated Cell Cultures (1 mentions)
Bafilomycin a1 ab120497, by Abcam (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Cisplatin, by Merck Group (1 mentions)
Mcf 7, by National Collection of Authenticated Cell Cultures (1 mentions)
Ana 12, by MedChemExpress (1 mentions)
Mcf 7 human breast cancer cell line, by National Collection of Authenticated Cell Cultures (1 mentions)
Ifosfamide, by Merck Group (1 mentions)
Ab154794, by Abcam (1 mentions)
Sc 397, by Santa Cruz Biotechnology (1 mentions)
A5441, by Merck Group (1 mentions)
6 protocols using adriamycin adm
1
Ovarian Cancer Cell Line Experiments
2
Culturing Breast Cancer Cell Lines
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Human breast cancer cell lines MCF-7, MDAMB231, Hs578t, and human embryonic kidney (HEK) 293T cells were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Human breast cancer multidrug resistance cell MCF-7/MDR was purchased from the Cell Bank of Xiangya Medical College, Central South University. Cells were cultured in Dulbecco’s Modified Eagle’s Medium (Gibco, California, USA) supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific, Beijing, China), 10 mmol/l HEPES, at 37 °C and 5% CO2. Adriamycin (ADM), 5-fluorouracil (5-FU), and cisplatin (DDP) were purchased from sigma (Sigma, MO, USA).
3
Breast Cancer Cell Line Cultivation
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The MCF-7 human breast cancer cell line was obtained from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). The multidrug-resistant human breast cancer cell MCF-7/MDR was purchased from the Cell Bank of Xiangya Medical College, Central South University (Changsha, China). Cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% (v/v) fetal bovine serum (FBS; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml streptomycin (Gibco; Thermo Fisher Scientific, Inc.) and 10 mmol/l HEPES at 37°C in a humidified atmosphere containing 5% CO2. Adriamycin (ADM), 5-fluorouracil (5-FU), cisplatin (DDP) and L-N5-(1-Iminoethyl)-ornithine hydrochloride (LNIO) were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). ANA-12 was purchased from MedChemExpress (Monmouth Junction, NJ, USA).
4
Investigating DNA Damage Response Pathways
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Cells were seeded at a density of 60%–70% and cultured for 24 hours before 12-hour drug treatments. DNA-damaging reagents, cisplatin (CDDP, Qiru Corp., People’s Republic of China), adriamycin (ADM, Sigma-Aldrich, St Louis, MO, USA), and ifosfamide (Sigma-Aldrich), were diluted to the indicated concentrations in cell culture medium. The PLK2 inhibitor ELN58264620 (link),21 (link) (Elan Pharmaceuticals, San Francisco, CA, USA) was diluted to 5 μg/mL in cell culture medium. Dimethyl sulfoxide was used as a control. The following antibodies were used for Western blot (WB) and coimmunoprecipitation (co-IP): anti-PLK2 (EPR10070, ab154794, Abcam, USA), anti-TAp73 (38C674.2, ab79078, Abcam, USA), anti-PUMA (4976S, CST, MA, USA), anti-p21 (SC-397, Santa Cruz Biotechnology, Inc., Dallas, TX, USA), anti-Flag (F3165, Sigma-Aldrich), and anti-β-actin (A5441, Sigma-Aldrich). The following antibodies were used for immunofluorescence assay: primary antibodies against PLK2 (EPR10070, ab154794, Abcam, MA, USA) and TAp73 (1E8, ab118985, Abcam, USA) and secondary antibodies against PLK2 (anti-rabbit 594, ZF-0416, ZSGB-BIO, People’s Republic of China) and TAp73 (anti-mouse-488, ZF-0412, ZSGB-BIO, People’s Republic of China).
5
Investigating SIRT1 Inhibition in Cancer
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Adriamycin (ADM) and 1-methylnicotinamide (MNA) were obtained from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO, USA), and paclitaxel (PTX) and the selectively SIRT1 inhibitor EX527 were obtained from Selleck Chemicals (SelleckChemicals, Houston, TX, USA). The anti-SIRT1, anti-β-actin, and anti-acetyl-p53 were all obtained from Cell Signaling Technology (CST, Beverly, MA, USA). The mouse anti-NNMT monoclonal antibody 1E7 was prepared through the hybridoma technique as previously described [14 (link)].
6
Breast Cancer Cell Line Manipulation
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MCF-7 (luminal A representative) and BT-474 (luminal B representative) cell lines were cultured as we previously described.11 (link) lentiviral MORC4 expression plasmid (MORC4-OE); lentiviral MORC4 expression plasmid with FLAG tag (MORC4-FLAG); lentiviral STAT3 expression plasmid with HA tag (STAT3-HA); lentiviral MID2 shRNA based on a validated inhibiting sequence from Sigma Aldrich (sh-MID2, 5ʹ-CGAGCGAATTGCTTGCCAATT-3ʹ); lentiviral MID2 expression plasmid with silent mutations of shRNA binding sites (MID2-MT-OE); MORC4 siRNAs (#1, 5ʹ-GCTTTGGCTTTACAGATAA-3ʹ; #2, 5ʹ-GCAAGAAACACATGGGTTA-3ʹ and #3, 5ʹ-CCATCCATGTTACCTGCAA-3ʹ), STAT3 siRNAs (#1, 5ʹ-GCAACAGATTGCCTGCATT-3ʹ; #2, 5ʹ-CCCGTCAACAAATTAAGAA-3ʹ and #3, 5ʹ-GGTACAACATGCTGACCAA-3ʹ) were produced by GeneChem Co., Ltd. (Shanghai, China). Empty lentiviral plasmids (Vector), vector carrying scramble shRNA (Sh-NC), and scramble siRNA were used as controls. Recombinant lentiviruses production was described previously.11 (link)
For si-RNA transfection, cells were co-transfected using either 20 pmol siRNA and 1 μg of different PGL3 promoter constructs (24-well plate) or 75 pmol siRNA only, using Lipofectamine 3000 (Invitrogen). Adriamycin (ADM), 5-fluorouracil (5-FU), and cisplatin (DDP) were purchased from Sigma-Aldrich (Sigma, MO, USA).
For si-RNA transfection, cells were co-transfected using either 20 pmol siRNA and 1 μg of different PGL3 promoter constructs (24-well plate) or 75 pmol siRNA only, using Lipofectamine 3000 (Invitrogen). Adriamycin (ADM), 5-fluorouracil (5-FU), and cisplatin (DDP) were purchased from Sigma-Aldrich (Sigma, MO, USA).
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