Flo 1

BAY1143572 was purchased from Active Biochemical (Wan Chai, Hong Kong). Human esophageal adenocarcinoma cell lines, FLO1, OE33, and SKGT4 were purchased from Sigma Aldrich or ATCC. Radiation resistant esophageal adenocarcinoma cells (OE33R) were provided by our institutional Center for Radiation Oncology Research. Complete cell line information is provided in the Supplementary Materials and Methods.

Human esophageal cancer cell lines (OE19, OE33, ESO26, KYSE270, SK-GT-2, Flo-1, ESO51, OE21, and OACM5.1C) were obtained from Sigma Aldrich (St. Lois, MO) and cultured according to manufacturer’s instructions. Flavopiridol was obtained from Cayman Chemical (Ann Arbor, MI), nanoparticle albumin-bound paclitaxel from Goshen Center for Cancer Care (Goshen, IN), BMS-754807 from Active Biochemical Limited (Maplewood, NJ), the cell proliferation reagent WST-1 from Roche Diagnostic Corporation (Indianapolis, IN), c-Myc siRNA (sc-29226) and control siRNA (sc-37007) from Santa Cruz Biotechnology (Santa Cruz, CA). pcDNA3-cMyc was a gift from Wafik El-Deiry (Addgene plasmid #16011) (Ricci et al., 2004 (link)) and pcDNA3-EGFP as a negative control (Addgene plasmid #13031) was purchased.

Esophageal adenocarcinoma cells OE33, FLO-1 and SKGT4 were purchased from Sigma-Aldrich (St. Louis, MO). ESO51 and KYAE-1 cells were obtained from Culture Collections (Public Health England, UK). OE33, ESO51 cells were maintained in a RPMI medium containing 2 mM of L-Glutamine, 10% of fetal bovine serum (FBS), 100 units/ml penicillin and 100 μg/ml of streptomycin. KYAE-1 cells were maintained in a medium of RPMI + Hams F12 (1:1) and FLO-1 and SKGT4 cells were maintained in a DMEM medium containing 10% of FBS, 100 units/ml penicillin and 100 μg/ml of streptomycin. 293 FT cells were obtained from Invitrogen (Carlsbad, CA) and maintained in a DMEM medium supplemented with 10 % FBS and 500μg/ml G418. Normal esophageal epithelial cells, HET-1A were provided by Dr. Xu (MD Anderson Cancer Center, Houston, TX) and maintained in a KSF medium (Lonza Walkersville Inc., Walkersville, MD). Cells were maintained in a 5% CO₂ atmosphere at 37°C and passaged at 80% confluence using 1 mM EDTA-0.025% trypsin for 3 to 5 minutes. All cell lines were authenticated by cell line validation core facility of UT M.D. Anderson Cancer Center.

Non-dysplastic BE and dysplastic esophageal cell lines CP-A, CP-B and CP-D were provided by Dr P. Rabinovitch (University of Washington, USA); BART-T cells were provided by Dr Rhonda Souza (Baylor University Medical Center, TX). All non-dysplastic BE and dysplastic esophageal cells were cultured in Keratinocyte Serum Free Medium (KSFM) in 2.5% heat inactivated fetal bovine serum (HI FBS) (Gibco), supplemented with 0.1 μg/mL Epidermal Growth Factor (PeproTech) and 0.7 μg/mL of Bovine Pituitary Extract (Sigma). OE33 and FLO-1 were purchased from the European Collection of Authenticated Cell Cultures (Sigma). OE33 were cultured in Roswell Park Memorial Institute 1640 medium (RPMI) in 10% HI FBS, while FLO-1 were cultured in Dulbecco’s modified Eagle’s medium (DMEM) in 2.5% HI FBS. All cell lines were cultured in a 5% CO2 incubator set to 37°C. FBS used throughout this study was heated to inactivate complement components.

EAC cell lines (FLO-1 and OE19) were purchased from Sigma Aldrich Corporation (Saint Louis, MO) and cultured as described previously [20 (link)-24 (link)].

Five EGJ adenocarcinoma cell lines, OACM5.1C, OE19, OE33, SK-GT-4 and FLO-1, were purchased from the European Collection of Cell Cultures (ECACC; Salisbury, UK). FLO-1 was cultured in DMEM (Sigma-Aldrich, St Louis, MO, USA) containing 10% fetal bovine serum (FBS) in a 5% CO₂ air-humidified atmosphere at 37°C. The other cell lines (OACM5.1C, OE19, OE33 and SK-GT-4) were cultured in RPMI-1640 (Life Technologies, Carlsbad, CA, USA) under the same conditions, according to the manufacturer's recommendations.