Celecoxib

Manufactured by Selleck Chemicals

Sourced in United States, China

Celecoxib is a chemical compound used as a laboratory reagent. It is a selective inhibitor of the cyclooxygenase-2 (COX-2) enzyme. Celecoxib is commonly used in research applications to study the role of COX-2 in various biological processes.

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Lab products found in correlation

43 protocols using celecoxib

Microglial Cell Culture and Treatments

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Mouse microglial BV2 cells (Cell Resource Center of the Chinese Academy of Medical Sciences) were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) with 10% FBS (Beijing Aoqing Biotechnology Co., Ltd.) and 1% penicillin/streptomycin at 37˚C in a humidified atmosphere with 5% CO₂ and the medium was changed every 2 days. Cells were split with 0.125% trypsin when they reached 80% confluence and the passages 2-10 were used for carrying out experiments. For LPS treatment, the cells were treated with 100 ng/ml LPS (cat. no. L2630; Sigma-Aldrich; Merck KGaA) for the indicated times. To perform the hypoxia exposure experiments, BV2 cells seeded in 60-mm dishes (8x10⁵ cells/dish) were put in the cell culture chamber with 1 or 3% oxygen (O₂) at 37˚C for the indicated times. For the FG-4592 (Selleck Chemicals) treatment, cells were treated with 10 µM FG-4592 for 6, 12 and 24 h at 37˚C. For the celecoxib (Selleck Chemicals) treatment, cells were pretreated with 10 and 20 µM celecoxib for 1 h at 37˚C.

Yang Y., Geng Y., Cheng X., Gao J., Shi Z, & Zhao M. (2023). Cyclooxygenase‑2 contributes to the hypoxia‑induced aggravation of the neuroinflammation response stimulated by lipopolysaccharide in microglia. Experimental and Therapeutic Medicine, 25(3), 123.

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Celecoxib and M. bovis Interaction

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Mice were randomly assigned to four experimental groups: (1) Control group (n = 7) (CT); (2) Celecoxib (Selleck, USA) +M. bovis group (n = 7) (Cele + M. bovis); (3) M. bovis infection group (n = 7) (M. bovis); (4) Celecoxib (20 μg/kg) (Selleck, USA) + Dimethyl PGE2 (50 μg/mL per mouse) +M. bovis group (n = 7) (Cele + dm-PGE2 + M. bovis). Celecoxib or dm-PGE2 was administered intraperitoneally for 5 weeks. The mice were sacrificed after 5 weeks of infection. The fresh stool was collected immediately from the colonic tissue of each mouse using sterile sampling containers (Supplement Figure 1B).

Wang H., Yao J., Chen Y., Wang Y., Liu Y., Liao Y., Liang Z., Dong Y.H., Qu M., Ge X, & Zhou X. (2022). Gut dysbacteriosis attenuates resistance to Mycobacterium bovis infection by decreasing cyclooxygenase 2 to inhibit endoplasmic reticulum stress. Emerging Microbes & Infections, 11(1), 1806-1818.

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Selective COX-2 Silencing in Cells

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COX-2 siRNA (sequence, 5′-GCTCAGCCATACAGCAAAT-3′) was purchased from Guangzhou RiboBio Co., Ltd. (Guangzhou, China). Cells were transfected Lipofectamine^® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol (siRNA, 100 nmol/l; 37°C for 24 h). The control siRNA was provided by Guangzhou RiboBio Co., Ltd. (Guangzhou, China) which was a random sequence of siRNA and incompatible with the target gene. Cells were treated with celecoxib (Selleck Chemicals, Houston, TX, USA) at the concentration of 1 mol/l (diluted in DMSO) at 37°C for 48 h (DMSO without celecoxib as the control).

Wang Y.X., Zhang Z.Y., Wang J.Q., Qian X.L, & Cui J. (2018). HPV16 E7 increases COX-2 expression and promotes the proliferation of breast cancer. Oncology Letters, 16(1), 317-325.

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Drug Screening and Combination Experiments

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Drugs used in this study: aprepitant (Selleck Chemicals, Cat# S1189), auranofin (Santa Cruz Biotechnology, Cat #sc-202476), captopril (Selleck Chemicals, Cat# S2051), celecoxib (Selleck Chemicals, Cat# S1261), copper(II)chloride dehydrate (Sigma-Aldrich, Cat# C3279), disulfiram (Selleck Chemicals, Cat# S1680), itraconazole (Selleck Chemicals, Cat# S2476), minocycline (Selleck Chemicals, Cat# S4226), quetiapine fumarate (Selleck Chemicals, Cat# S1763), sertraline (Selleck Chemicals, Cat# S4052), and temozolomide (Sigma-Aldrich, Cat# T2577). copper(II)chloride dehydrate (CuCl₂) was added to all wells containing disulfiram (DSF) and corresponding control wells (Skrott et al. 2017 (link)). A fixed concentration of 20 µM Cu was used in this study (Twomey et al. 2008 (link)). minocycline was dissolved in H₂O, while all other drugs were dissolved in DMSO for generation of stock solutions and stored according to the manufacturer’s instructions.

Skaga E., Skaga I.Ø., Grieg Z., Sandberg C.J., Langmoen I.A, & Vik-Mo E.O. (2019). The efficacy of a coordinated pharmacological blockade in glioblastoma stem cells with nine repurposed drugs using the CUSP9 strategy. Journal of Cancer Research and Clinical Oncology, 145(6), 1495-1507.

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Celecoxib and Radiation-Induced Apoptosis

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Celecoxib was purchased from Selleck Chemicals (Houston, TX, USA). X-ray radiation was conferred by Radsource 2000 from Radsource, LLC (Brentwood, TN, USA). TRIzol^® and primers were purchased from Invitrogen (Thermo Fisher Scientific, Inc., Waltham, MA, USA) and the ThermoScript RT reverse transcription-polymerase chain reaction (RT-PCR) kit was purchased from Fermentas (Thermo Fisher Scientific, Inc.). The PCR Amplification kit was obtained from Takara Bio, Inc., (Otsu, Japan). The Annexin V-Fluorescein Isothiocyanate (FITC) kit (cat. no. KFG001) was purchased from Nanjing KeyGen Biotech Co., Ltd. (Nanjing, China). Hoechst 33258 was purchased from Beyotime Institute of Biotechnology (Haimen, China). The laser confocal scanning microscope, flow cytometer, PCR thermocycler, gel electrophoresis imaging system and cell culturing equipment were all obtained from The Second Affiliated Hospital, Suzhou University (Suzhou, China).

Han Z.Q., Liao H., Shi F., Chen X.P., Hu H.C., Tian M.Q., Wang L.H, & Ying S. (2017). Inhibition of cyclooxygenase-2 sensitizes lung cancer cells to radiation-induced apoptosis. Oncology Letters, 14(5), 5959-5965.

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NPC Cell Lines and Pharmacological Agents

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Human NPC cell lines (CNE1 and CNE2) were from the Cancer Center of Sun Yat-sen University (Guangzhou, China). Cells are maintained in Dulbecco's Modified Eagle Medium (DMEM, Gibco, NY, USA) with 10% fetal bovine serum (FBS, Gibco, CA, USA) and 1% penicillin-streptomycin (Gibco) at 37°C in a 5% CO₂ humidified incubator (Thermo, CO, USA). Hoechst 33342, dimethyl sulfoxide (DMSO), verapamil, RSV, Mdivi-1, 5-FU were purchased from Sigma (MO, USA). Aspirin, celecoxib and indomethacin were purchased from Selleck (TX, USA).

Zhou T.J., Zhang S.L., He C.Y., Zhuang Q.Y., Han P.Y., Jiang S.W., Yao H., Huang Y.J., Ling W.H., Lin Y.C, & Lin Z.N. (2017). Downregulation of mitochondrial cyclooxygenase-2 inhibits the stemness of nasopharyngeal carcinoma by decreasing the activity of dynamin-related protein 1. Theranostics, 7(5), 1389-1406.

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Fluorescent Screening of COX Inhibitors

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Cyclooxygenase inhibition was determined using the COX Fluorescent Inhibitor Screening Assay Kit (Cayman Chemical) according to the manufacturer’s recommendation. SS (Sigma-Aldrich) (10 µM) served as a positive control for assessing the COX-1 and COX-2 inhibitory activity of ADT 061. Celecoxib (Selleck Chemicals) (100 nM) and indomethacin (Sigma-Aldrich) (100 nM) served as additional controls as COX-2 and COX-1 specific inhibitors, respectively. The protocol was modified to include a 20-minute incubation step prior to the addition of arachidonic acid. Fluorescent readings were obtained using a Biotek Synergy H4 plate reader.

Lee K.J., Chang W.C., Chen X., Valiyaveettil J., Ramirez-Alcantara V., Gavin E., Musiyenko A., da Silva L.M., Annamdevula N.S., Leavesley S.J., Ward A., Mattox T., Lindsey A.S., Andrews J., Zhu B., Wood C., Neese A., Nguyen A., Berry K., Maxuitenko Y., Moyer M.P., Nurmemmedov E., Gorman G., Coward L., Zhou G., Keeton A.B., Cooper H.S., Clapper M.L, & Piazza G.A. (2021). Suppression of colon tumorigenesis in mutant Apc mice by a novel PDE10 inhibitor that reduces oncogenic β-catenin. Cancer prevention research (Philadelphia, Pa.), 14(11), 995-1008.

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Evaluating β-catenin Inhibitor iCRT14 in Mice

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After surgery, all mice were allowed to rest for 4 weeks and then treated with the β-catenin inhibitor iCRT14 for 6 weeks. iCRT-14 (S8704) was purchased from Selleck (TX, USA) and was first dissolved in DMSO and then further dissolved in corn oil (50 mg·kg⁻¹ body weight) and administered every 3 days for 6 weeks by i.p. injection. Injection of corn oil (S6701, Selleck, USA) was used as a negative control, and celecoxib (S1261, Selleck, USA) was used as a positive control. celecoxib was also first dissolved in DMSO and then dissolved in corn oil (10 mg·kg⁻¹ body weight) and delivered by i.p. injection on the same schedule as iCRT-14.

Lu K., Wang Q., Jiang H., Li J., Yao Z., Huang Y., Chen J., Zhang Y., Xiao G., Hu X., Luo Z., Yang L., Tong L, & Chen D. (2023). Upregulation of β-catenin signaling represents a single common pathway leading to the various phenotypes of spinal degeneration and pain. Bone Research, 11, 18.

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Effects of MIF, COX-2 on Cell Growth

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BPH-1 cells and PWR-1E cells were plated on 96-well plates (1000 cells per well) overnight and rMIF (300-69, PeproTech, USA; 100 ng/ml), MIF inhibitor ISO-1 (S7732, Selleck, USA; 10 µM), COX-2 inhibitor celecoxib (S1261, Selleck, USA; 5 µM) or nothing was added into each well. BPH-1 and PWR-1E cell growth was assessed on days 2, 4, and 6 using the Cell Counting Kit-8 (CCK8) (CK04; Dojindo Molecular Technologies, Tokyo, Japan) according to the manufacturer's protocol. The absorbance value was measured using a Varioskan Flash plate reader (Thermo Fisher Scientific, Waltham, MA, USA) at 450 nm.

Song H., Shen Q., Hu S, & Jin J. (2020). The role of macrophage migration inhibitory factor in promoting benign prostatic hyperplasia epithelial cell growth by modulating COX-2 and P53 signaling. Biology Open, 9(11), bio053447.

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PET/MRI Imaging of Tumor-Bearing Mice

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PET/MR image acquisition and analysis were performed as previously described [9 (link)]. Tumor-bearing mice were anesthetized by isoflurane in O₂ (4 %, 2 l/min induction; 1–2 %, 1 l/min maintenance) for lateral tail-vein catheterization then transferred to a nanoScan™ PET/MRI 3T scanner (Mediso). Mice bearing HT-29 or HCT-116 xenografts (n = 5) were injected through the tail-vein catheter with [¹¹C]MC1 (4.39–13.35 MBq, 0.47–10.81 nmol/kg, 39–509 GBq/μmol). HT-29 xenograft mice were pre-treated with an intraperitoneal injection of 2 mg in 20 μl dimethylsulfoxide of unlabeled MC1 (n = 2) or the known COX-2 inhibitor, celecoxib (n = 3; Selleck Chemicals), 60 min prior to injection of [¹¹C]MC1. Following the 60 min PET scans, mice were sacrificed by cervical dislocation and tissue samples were collected, weighed, and transferred to γ-counting tubes for biodistribution analysis. Tissue radioactivity was measured with a γ-counter and expressed as %ID/g. Image analyses and extraction of time-activity curves (TACs) from regions of interest (ROIs) were performed in Amide v1.0.4.

Boyle A.J., Narvaez A., Tong J., Zoghbi S.S., Pike V.W., Innis R.B, & Vasdev N. (2021). Repurposing [11C]MC1 for PET Imaging of Cyclooxygenase-2 in Colorectal Cancer Xenograft Mouse Models. Molecular imaging and biology, 24(3), 365-370.

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