Gemcitabine

Manufactured by Thermo Fisher Scientific

Sourced in United States

Gemcitabine is a laboratory reagent used in scientific research. It is a synthetic pyrimidine nucleoside analog that acts as an antimetabolite, inhibiting DNA synthesis and cell growth. Gemcitabine is commonly used in studies involving cell biology, molecular biology, and oncology research.

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Lab products found in correlation

Cisplatin, by Merck Group (2 mentions) Gemcitabine, by Merck Group (2 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Doxorubicin, by Thermo Fisher Scientific (1 mentions) Vincristine, by Thermo Fisher Scientific (1 mentions) Deferoxamine mesylate, by Thermo Fisher Scientific (1 mentions) Celltiter glo 2, by Promega (1 mentions) Sn 38, by Selleck Chemicals (1 mentions) Ku 60019, by Selleck Chemicals (1 mentions) Sorafenib, by Thermo Fisher Scientific (1 mentions) Prism software, by GraphPad (1 mentions) Celecoxib, by Merck Group (1 mentions) Rmp1 14, by BioXCell (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Trifluoroacetic acid, by Thermo Fisher Scientific (1 mentions) Hyamine hydroxide, by PerkinElmer (1 mentions) Thymidine, by Thermo Fisher Scientific (1 mentions) Cytoscint scintillation solution, by MP Biomedicals (1 mentions) Cell culture reagents and hemoglobin standard, by Merck Group (1 mentions)

6 protocols using gemcitabine

Bladder cancer cell culture

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Human bladder epithelial immortalized cell line (SV-HUC-1) and BUC cell lines (J82, T24) were purchased from the American Type Culture Collection (Manassas, USA). Those Cells were cultured in Dulbecco's modified Eagle's medium (DMEM) containing with 10% fetal bovine serum (Thermo Fisher Scientific, USA) in a 95% air/5% CO₂ incubator under 37°C.
Gemcitabine-resistant T24/Gem cell line was set up previously through induction with low concentration of Gemcitabine in our laboratory. To maintain the resistant phenotype, T24/Gem cells was cultured with 0.5 μg/mL Gemcitabine (Sigma, USA), and cultured in Gemcitabine-free medium for 7 days prior to experiment.

Xie D., Zhang H, & Shang C. (2018). Long non-coding RNA CDKN2B antisense RNA 1 gene inhibits Gemcitabine sensitivity in bladder urothelial carcinoma. Journal of Cancer, 9(12), 2160-2166.

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High-throughput Drug Screening in HepG2 Cells

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HepG2 or HB214 (1500 cells/well) was seeded on a 384 well plate in 30 μl. Drugs were added after 24 h. Cell Titer Glo (CTG) viability assay was conducted after 72 h of drug treatment. Briefly, CellTiter-Glo 2.0 (#G9243 Promega) was added to each well in a 1:1 v/v ratio. Plates were then covered to keep from light and incubated at RT on an orbital shaker at 150 RPM for 30 mins. After the incubation, plate was read on a synergy H4 plate reader for luminescence. Dose effect curves for each drug were calculated using Prism software, version 9 (GraphPad). For drug combinations, responses were analyzed using SynergyFinder2.0^{50 (link)}. Drugs used include cisplatin (#479036-5 G, Sigma Aldrich), gemcitabine (#AC456890010, Fisher Scientific), vincristine (#AAJ60907MA, Fisher Scientific), triapine (#50-136-4826, Fisher Scientific), MK1775 (#M4102, LKT laboratories, Saint Paul, Minnesota), doxorubicin (#BP25161, Fisher Scientific), sorafenib (#NC0749948, Fisher Scientific), SN-38 (#S4908-50MG, Selleck Chemicals, Harris County, Texas), deferoxamine mesylate (#AC461770010, Acros Organics, Geel, Belgium), KU60019 (#S1570, Selleck Chemicals). All concentrations were seeded in triplicate and the experiment was repeated three times. Significance was determined using the Extra Sum of Squares f test.

Brown A., Pan Q., Fan L., Indersie E., Tian C., Timchenko N., Li L., Hansen B.S., Tan H., Lu M., Peng J., Pruett-Miller S.M., Yu J., Cairo S, & Zhu L. (2023). Ribonucleotide reductase subunit switching in hepatoblastoma drug response and relapse. Communications Biology, 6, 249.

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Combination Immunotherapy for Tumor Regression

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Mice with established G69 or G7 tumors (i.e., tumor volume of ~50 mm³) were randomized into treatment groups (using RANDOM.com) and injected intraperitoneally (i.p.) with either vehicle (10% DMSO in 0.05% NaCl solution) or GC (gemcitabine at 120 mg/Kg; Fisher Scientific and cisplatin at 6 mg/Kg; Sigma–Aldrich) on day one and gemcitabine on day 8 to complete one cycle of chemotherapy. Mice were left to recover from chemotherapy for seven days upon which they received the second cycle of GC chemotherapy. Two days after the last gemcitabine treatment, tumors were harvested and processed for subsequent analyses. For the iDAMP blockade group, mice received celecoxib (10 mg/Kg; Sigma–Aldrich) daily until the completion of the experiment. For single therapy treatment, anti-PD1 antibody (200 ug/mouse, RMP1-14, Bio x Cell) was injected i.p. every other day for a total of three doses. For combination treatments, anti-PD1 was injected i.p. on days 9, 11, 13 of each chemotherapy cycle.

Nikolos F., Hayashi K., Hoi X.P., Alonzo M.E., Mo Q., Kasabyan A., Furuya H., Trepel J., Di Vizio D., Guarnerio J., Theodorescu D., Rosser C., Apolo A., Galsky M, & Chan K.S. (2022). Cell death-induced immunogenicity enhances chemoimmunotherapeutic response by converting immune-excluded into T-cell inflamed bladder tumors. Nature Communications, 13, 1487.

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Establishment of Drug-Resistant MSTO-211H Cell Lines

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The human MPM cell line MSTO-211H was obtained from the ATCC. The parental cells (MSTO-Par) and sub-lines resistant to cisplatin (MSTO-CisR), gemcitabine (MSTO-GemR) and vinorelbine (MSTO-VinoR) were cultured in RPMI-1640 medium supplemented with 10% FCS (both Thermo Fisher Scientific, Carlsbad, CA, USA) with 5% CO₂ at 37 °C. Drug-resistant cell lines (MSTO-CisR, MSTO-GemR and MSTO-VinoR) were established by standard cyclic treatment with the IC₅₀ of cisplatin, gemcitabine and vinorelbine (all drugs purchased from Sigma Aldrich, St. Louis, MO), respectively. Drug concentrations were increased incrementally until a resistance of 2 to 5-fold was reached, after which they were maintained by treatment with constant concentrations of drug (10 µmol/L for cisplatin and 20 nmol/L for gemcitabine and vinorelbine) from 24 h following passaging. Resistance gradually increased over a period of 3-4 months to the final resistance levels described in Results. Resistance was regularly confirmed by comparison of drug IC₅₀ values in the drug resistant cell lines and parental line.

Williams M., Cheng Y.Y., Phimmachanh M., Winata P., van Zandwijk N, & Reid G. (2019). Tumour suppressor microRNAs contribute to drug resistance in malignant pleural mesothelioma by targeting anti-apoptotic pathways. Cancer Drug Resistance, 2(4), 1193-1206.

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Radiolabeled Gemcitabine Metabolism Analysis

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Gemcitabine, dilazep hydrochloride, thymidine, and high-performance liquid chromatography (HPLC) grade acetonitrile and trifluoroacetic acid (TFA) were purchased from Thermo Fisher Scientific (Waltham, MA). The deaminated Gemcitabine metabolite dFdU was purchased from Sigma-Aldrich (St. Louis, MO). Radiolabeled Gemcitabine (cytosine-2-¹⁴C) (55.0 mCi/mmol), subsequently referred to as [¹⁴C]-Gemcitabine, was purchased from Moravek, Inc. (Brea, CA). CytoScint™ scintillation solution was purchased from MP Biomedicals, LLC (Solon, OH). Hyamine hydroxide was purchased from PerkinElmer (Waltham, MA). All other chemicals were obtained from standard commercial sources.

Thompson B.R., Hu Y, & Smith D.E. (2019). Mechanisms of Gemcitabine Oral Absorption as Determined by In Situ Intestinal Perfusions in Mice. Biochemical pharmacology, 168, 57-64.

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Investigating Pancreatic Cancer Progression

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Human pancreatic cancer cell lines, MPanc96 and SUIT2 expressing firefly luciferase, were provided by Dr. Arumugam (MD Anderson Cancer Center, Houston, TX). Cell culture reagents and hemoglobin standard, Drabkin’s reagent, and other common reagents were purchased from Sigma (St. Louis, MO). D-Luciferin potassium salt was purchased from Caliper Life Sciences (Hopkinton, MA), and gemcitabine was purchased from Thermo Fisher Scientific (Waltham, MA). Matrigel was purchased from BD Bioscience (San Jose, CA). Tinzaparin was obtained from Leo Pharma Inc. (Ballerup, Denmark), and S-NACH was synthesized at Rensselaer Polytechnic Institute (Rensselaer, NY). Anti-pSer15-p53 (9286) was obtained from Cell Signaling (Danvers, MA), Anti-XIAP (sc-55550), Anti-THBS1 (sc-65612), and Anti-p21 (sc-6246) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX).

Sudha T., Yalcin M., Lin H.Y., Elmetwally A.M., Nazeer T., Arumugam T., Phillips P, & Mousa S.A. (2014). Suppression of Pancreatic Cancer by Sulfated Non-Anticoagulant Low Molecular Weight Heparin. Cancer letters, 350(0), 25-33.

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