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Etoposide

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Etoposide is a topoisomerase II inhibitor used in laboratory research. It functions by interfering with the activity of the enzyme topoisomerase II, which is involved in DNA replication and transcription. Etoposide is commonly used in cell biology and cancer research studies.

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Lab products found in correlation

Phosphoric acid, by Fujifilm (1 mentions) Camptothecin, by TopoGEN (1 mentions) Paclitaxel, by Selleck Chemicals (1 mentions) Supercoiled phot1 dna, by TopoGEN (1 mentions) Dimethylacetamide dma, by Merck Group (1 mentions) Cremophor el, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Matrigel, by BD (1 mentions) Pericentrin, by Abcam (1 mentions) Chloroquine, by Thermo Fisher Scientific (1 mentions) Topo 2 assay kit, by TopoGEN (1 mentions) Ab4448, by Abcam (1 mentions) 17 aag, by Thermo Fisher Scientific (1 mentions) Doxorubicin, by Thermo Fisher Scientific (1 mentions) Bi2536, by Selleck Chemicals (1 mentions) Ab6276, by Abcam (1 mentions)

3 protocols using etoposide

1

Synthesis and Procurement of Anticancer Agents

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HH-N25 was synthesized through an established protocol in our lab,39 (link) while paclitaxel was procured from Selleckchem (Houston, TX, USA). TOPI and TOPII, supercoiled pRYG DNA, supercoiled pHOT1 DNA, camptothecin, and etoposide were purchased from TopoGEN. All other reagents and chemicals including DMEM (Gibco-Invitrogen, Grand Island, NY, USA), Matrigel (BD Biosciences, USA), Cremophor EL (Sigma, St. Louis, MO, USA), FBS (Hyclone, Logan, UT, USA), dimethylacetamide (DMA; Sigma), estradiol cyclopentyl propionate (Estol-depot injection, Astar, Taipei City, Taiwan), and phosphoric acid (Wako, Japan) were procured from reputable manufacturers.
Lawal B., Kuo Y.C., Sumitra M.R., Wu A.T, & Huang H.S. (2021). In vivo Pharmacokinetic and Anticancer Studies of HH-N25, a Selective Inhibitor of Topoisomerase I, and Hormonal Signaling for Treating Breast Cancer. Journal of Inflammation Research, 14, 4901-4913.
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2

DNA Topoisomerase II Binding Assay

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DPBQ and other chemicals were obtained from the National Cancer Institute, diluted in stock DMSO solutions and used at the concentrations shown. Other chemicals used include doxorubicin (ThermoFisher, Waltham MA), etoposide (Topogen, Port Orange, FL), paclitaxel (ThermoFisher/Acros), BI-2536 (Selleck Chemical, Houston, TX), chloroquine, 17-AAG, and Nutlin-3a (Fisher). Antibodies used include β-actin (ab6276 WB 1:10,000; Abcam,Cambridge, MA), pericentrin (ab4448 IF 1:1000; Abcam), p53 (#9282, WB 1:2000; Cell Signaling Technology, Beverly, MA), phospho-P15 p53 (#9284 WB 1:2000; Cell Signaling Technology), p21 (sc6246, WB 1:500; Santa Cruz, Dallas, TX)
The topoisomerase assay was performed using the Topo II Assay kit per the manufacturer instructions (TopoGEN, Port Orange, FL). Circular dichroism of 0.5 mg/ml salmon sperm DNA was performed using an AVIV Model 420 CD Spectrometer at room temperature. To analyze DNA binding, 100 μM ethidium bromide or DPBQ was added to the sample to evaluate changes in spectrum consistent with intercalation.
Choudhary A., Zachek B., Lera R.F., Zasadil L.M., Lasek A., Denu R.A., Kim H., Kanugh C., Laffin J.J., Harter J.M., Wisinski K.B., Saha S., Weaver B.A, & Burkard M.E. (2015). Identification of selective lead compounds for treatment of high-ploidy breast cancer. Molecular cancer therapeutics, 15(1), 48-59.
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3

Topoisomerase II-Mediated DNA Relaxation Assay

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The assay was performed following the manufacturer’s protocol, using a standard relaxation reaction mix (20 μL) containing 50 mM Tris-HCl (pH 8.0), 10 mM MgCl, 200 mM potassium glutamate, 10 mM dithiothreitol, 50 μg/mL bovine serum albumin, 1 mM ATP, 0.3 μg pHOT1 plasmid DNA, 8 units of human topo II (Topogen, Columbus, OH, USA), and etoposide (10 mM) or various concentrations of Apl-1, at 37 °C for 30 min. The reaction was terminated by adding 2 μL of 10% SDS, followed by 2.5 μL proteinase K (50 μg/mL) to digest the bound protein. The system was incubated at 37 °C for 15 min. The DNA product was finally analyzed by electrophoresis on a vertical 2% agarose gel at 2 volts/cm in 0.5× TAE buffer, and photographed using the Eagle Eye II system (Stratagene, La Jolla, CA, USA).
Shih S.P., Lu M.C., El-Shazly M., Lin Y.H., Chen C.L., Yu S.S, & Liu Y.C. (2022). The Antileukemic and Anti-Prostatic Effect of Aeroplysinin-1 Is Mediated through ROS-Induced Apoptosis via NOX Activation and Inhibition of HIF-1a Activity. Life, 12(5), 687.
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