The human pancreatic cancer cell lines MiaPaPc-2, BxPC-3 and PANC-1 were purchased from the American Type Culture Collection. The S2VP10 cells, derived from the Suit-2 cells as reported30 (link), were originally provided by Dr. M. Hollingsworth (University of Nebraska). Cell line authentication was determined by short tandem repeat analysis (STR) performed by the American Type Culture Collection. OGT overexpression MiaPaPc-2 and BxPC-3 cells were generated by transfection of pCMV-flag-OGT (OGT, Addgene, #29760); and OGT knockdown S2VP10 and PANC-1 cells were generated by infected with lentiviruses containing a short hairpin RNA (shRNA) specific for OGT (shOGT, Addgene, #10878). PANC-1 cells with FADD knockdown were established as we previously described by shRNA targeting FADD (Open Biosystems)31 (link). All stable clones were selected by 2 μg/ml puromycin as we previously reported31 (link). Sequencing analysis was performed to confirm the correct sequence, and protein expression was validated by Western blot analysis. Apoptosis was induced by TRA-8, a DR5 agonist antibody that was generated as previously described32 (link); and determined by Annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI) staining kit (BD Biosciences, San Jose, CA) and analyzed by flow cytometry as we previously reported11 (link).
Panc 1
Manufactured by Addgene
Sourced in United States
PANC-1 is a human pancreatic carcinoma cell line derived from an epithelioid carcinoma of the pancreas. It is a widely used in vitro model for pancreatic cancer research.
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Lab products found in correlation
Bxpc 3, by American Type Culture Collection (1 mentions)
Panc 1, by American Type Culture Collection (1 mentions)
Annexin 5 fluoresce in isothiocyanate fitc and propidium iodide staining kit, by BD (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Ptflc3, by Addgene (1 mentions)
2 protocols using panc 1
1
Modulation of Pancreatic Cancer Cell Apoptosis
2
Pancreatic Cell Lines and Islet Isolation
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Human pancreatic cell lines PANC-1 and pancreatic β-cell lines INS-1 were purchased from The Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). Islets were isolated from healthy rats and model rats and dispersed into single cells according to an earlier report [38 (link)]. PANC-1 cells were cultured in DMEM, and INS-1 and isolated islet cells were cultured in RPMI 1640 with 10% (v/v) FCS, 10 mM HEPES and 2 mM glutamine, 100 μg/mL penicillin, and 100 μg/mL streptomycin at 37°C and 5% CO2. Pancreatic cancer cell line PANC-1 was transfected with the plasmids ptfLC3 (an indicator of autophagic activity, Addgene; ID 21074, Cambridge, MA, USA), pcDNA3.1-NF-κB, and pTZU6+1-shRNA-NF-κB. Transfection was performed via Lipofectamine 2000TM (Invitrogen, Carlsbad, CA, USA). After a two-day transfection, the antibiotic-resistant cells were collected by using 200 μg/mL G-418. The strain was cultured at the same condition for three days. 25 mg/L picroside II was added to PANC-1, INS-1, and isolated islet cells and cultured in DMEM at 37°C and 5% CO2.
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