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Docetaxel

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Docetaxel is a chemotherapeutic agent used in the treatment of various types of cancer. It is a microtubule-stabilizing drug that inhibits cell division. Docetaxel is commonly used in the management of breast, lung, prostate, and other solid tumors.

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Lab products found in correlation

Docetaxel, by Sartorius (2 mentions) Incucyte zoom, by Sartorius (2 mentions) Powerplex 16 str system, by Promega (1 mentions) Pd98059, by Cayman Chemical (1 mentions) Gemcitabine, by Bio-Techne (1 mentions) Rerf lc ai, by RIKEN BioResource Center (1 mentions) Rerf lc kj, by RIKEN BioResource Center (1 mentions) Caffeine, by Bio-Techne (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) N acetyl l cysteine nac, by Merck Group (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Vinorelbine, by Santa Cruz Biotechnology (1 mentions) Pemetrexed, by Santa Cruz Biotechnology (1 mentions) Ku55933, by Bio-Techne (1 mentions) Gefitinib, by Cayman Chemical (1 mentions) Lc 2 ad, by RIKEN BioResource Center (1 mentions) Ly294002, by Cayman Chemical (1 mentions) A549 cells, by RIKEN BioResource Center (1 mentions) P iκb, by Cell Signaling Technology (1 mentions)

5 protocols using docetaxel

1

Comprehensive Cell Line Characterization

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The human NSCLC cell lines RERF-LC-AI, RERF-LC-KJ, and LC2/Ad were obtained from Riken BRC through the National Bio-Resource Project of the MEXT (Tsukuba, Japan). The A549 cells were purchased from both Riken BRC and American Type Culture Collection (Manassas, VA), while the PC–9 cell line was obtained from IBL cell bank (Gunma, Japan). The genotypes of all cell lines were identified with STR Identifier (Applied Biosystems) or PowerPlex 16 STR system (Promega). All the cell lines were maintained in culture in RPMI 1640 medium with 2mM L-glutamine (Invitrogen) supplemented with 10% FBS (Sigma-Aldrich) (A549, RERF-LC-AI, RERF-LC-KJ and PC–9) or 15% FBS (LC2/Ad) and 50 U/ml penicillin streptomycin (Sigma-Aldrich) at 37°C in a humidified atmosphere with 5% CO2. For cell culture work, Gefitinib (Cayman, #13166), Docetaxel (Enzo Life Sciences, BML-T129), LY294002 (Cayman, #70920), PD98059 (Cayman, #10006726) and KU55933 (Tocris Bioscience, #3544) were dissolved in DMSO (Sigma-Aldrich). Gemcitabine (Tocris Bioscience, #3259), Pemetrexed (Santa Cruz, #sc–219564), Vinorelbine (Santa Cruz, #sc–216059), Caffeine (Tocris Bioscience, #2793) and N-acetyl-L-cysteine (NAC) (Sigma-Aldrich, #A9165) were dissolved in PBS (-).
Okita R., Wolf D., Yasuda K., Maeda A., Yukawa T., Saisho S., Shimizu K., Yamaguchi Y., Oka M., Nakayama E., Lundqvist A., Kiessling R., Seliger B, & Nakata M. (2015). Contrasting Effects of the Cytotoxic Anticancer Drug Gemcitabine and the EGFR Tyrosine Kinase Inhibitor Gefitinib on NK Cell-Mediated Cytotoxicity via Regulation of NKG2D Ligand in Non-Small-Cell Lung Cancer Cells. PLoS ONE, 10(10), e0139809.
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2

Docetaxel Sensitivity Following SNAI1 Knockdown

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To assess changes in drug sensitivity following SNAI1 knockdown, stable cells (1,500 per well) were seeded in 96-well plates in the complete medium a day before treatment. A 2 mM stock solution of docetaxel (ENZO, New York, USA) was prepared in DMSO and was further diluted in complete medium to obtain eight serial dilutions such that the final treatment concentrations ranged from 0–2 μM. SNAIL-KD cells and the negative control (scrambled shRNA) cells were treated with DMSO or the various docetaxel dilutions and cell viability was determined using live imaging (Incucyte Zoom, Essen Biosciences). Phase contrast images (4x objective) were recorded at 0 hr, 16 hr and 24 hr after treatment initiation and the percentage confluence (a measure of cell viability) was assessed using the associated software as per manufacturer’s instructions. Relative viability was normalized to the confluence value treated with DSMO. docetaxel treatment was repeated independently to ensure reproducibility of the results. The Student’s t test was used to analyze differences, and p < 0.05 was considered statistically significant.
Peng X., Chen Z., Farshidfar F., Xu X., Lorenzi P.L., Wang Y., Cheng F., Tan L., Mojumdar K., Du D., Ge Z., Li J., Thomas G.V., Birsoy K., Liu L., Zhang H., Zhao Z., Marchand C., Weinstein J.N., Bathe O.F, & Liang H. (2018). Molecular Characterization and Clinical Relevance of Metabolic Expression Subtypes in Human Cancers. Cell reports, 23(1), 255-269.e4.
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3

Investigating Inflammatory Signaling Pathways

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Commercial antibodies against IL6, CXCL8, p65 (GeneTex International Corp, Hsinchu City, Taiwan), p105/p50, p100/52 (EMD Millipore, Billerica, MA), pIKKα/β, pIκB (Cell Signaling Technology, Danvers, MA), IκB (Santa Cruz, Dallas, Texas) and CD31, phosphorylated p65, IKKβ, IKKα and pSTAT3 (Abcam, Cambridge, MA) were used. Chemokine ELISA kits were from R&D systems (Minneapolis, MN). Docetaxel was from Enzo Life Sciences (Farmingdale, NY). Fluorescent dye-conjugated antibodies used in the flow cytometry analysis were all from Biolegend (San Diego, CA).
Tsai C.H., Tzeng S.F., Hsieh S.C., Yang Y.C., Hsiao Y.W., Tsai M.H, & Hsiao P.W. (2017). A standardized herbal extract mitigates tumor inflammation and augments chemotherapy effect of docetaxel in prostate cancer. Scientific Reports, 7, 15624.
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4

Preparation of Chemotherapy Stock Solutions

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A stock solution of cisplatin (Sigma) was dissolved in PBS to a concentration of 1.7 mmol/L (0.5 mg/mL), a stock solution of docetaxel (Enzo Life Sciences) was dissolved in DMSO to a concentration of 10 μmol/L (8 μg/mL), and a stock solution of gemcitabine (Gimzar, Lilly) was dissolved in PBS to a concentration of 10 mmol/L (3 mg/mL). All working solutions were freshly prepared in cell culture medium to the needed concentrations.
Wang Y., Gable T., Ma M.Z., Clark D., Zhao J., Zhang Y., Liu W., Mao L, & Mei Y. (2017). A piRNA-like Small RNA Induces Chemoresistance to Cisplatin-Based Therapy by Inhibiting Apoptosis in Lung Squamous Cell Carcinoma. Molecular Therapy. Nucleic Acids, 6, 269-278.
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5

Docetaxel Sensitivity Following SNAI1 Knockdown

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To assess changes in drug sensitivity following SNAI1 knockdown, stable cells (1,500 per well) were seeded in 96-well plates in the complete medium a day before treatment. A 2 mM stock solution of docetaxel (ENZO, New York, USA) was prepared in DMSO and was further diluted in complete medium to obtain eight serial dilutions such that the final treatment concentrations ranged from 0–2 μM. SNAIL-KD cells and the negative control (scrambled shRNA) cells were treated with DMSO or the various docetaxel dilutions and cell viability was determined using live imaging (Incucyte Zoom, Essen Biosciences). Phase contrast images (4x objective) were recorded at 0 hr, 16 hr and 24 hr after treatment initiation and the percentage confluence (a measure of cell viability) was assessed using the associated software as per manufacturer’s instructions. Relative viability was normalized to the confluence value treated with DSMO. docetaxel treatment was repeated independently to ensure reproducibility of the results. The Student’s t test was used to analyze differences, and p < 0.05 was considered statistically significant.
Peng X., Chen Z., Farshidfar F., Xu X., Lorenzi P.L., Wang Y., Cheng F., Tan L., Mojumdar K., Du D., Ge Z., Li J., Thomas G.V., Birsoy K., Liu L., Zhang H., Zhao Z., Marchand C., Weinstein J.N., Bathe O.F, & Liang H. (2018). Molecular Characterization and Clinical Relevance of Metabolic Expression Subtypes in Human Cancers. Cell reports, 23(1), 255-269.e4.
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