Hpde6 c7

Human pancreatic cancer cell lines including BXPC3, PANC-1, and ASPC-1 as well as human normal pancreatic cells HPDE6-C7 were obtained from the Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy. PANC-1, ASPC-1, and HPDE6-C7 were cultured in DMEM (sigma, D5796) with 10% fetal bovine serum (FBS), and 1% penicillin, streptomycin, and BXPC3 were cultured in RPMI-1640 medium (sigma, R8758) with 10% FBS and 1% penicillin and streptomycin at 37°C, 5% CO₂.

The normal human pancreatic ductal epithelial cell line (HPDE6-C7) was purchased from the Beijing Beinachuanglian Biotechnology Research Institute (Beijing, China) and cultured in an appropriate density with the recommended medium, supplemented with 10% fetal bovine serum in a humidified incubator containing 5% CO₂ at 37°C.
An LSR shRNA (targeting sequences of 5′-TGCTGAGCTACTCCTGTCAACGTCTCGTTTTGGCCACTGACTGACGAGACGTTCAGGAGTAGCT-3′) was cloned into a pcDNA6.2-GW/EmGFP-miR (Invitrogen, Carlsbad, CA, USA). This expression vector used plenti6.3/V5 DEST vectors. After amplification and DNA sequencing confirmation, this vector or negative control (NC) vector was used to generate lentiviruses in HEK-293 cells. These lentiviruses were used to infect HPDE6-C7 cells and named as LSR-knockdown (LSR-KD) cells or NC cells. In addition, a lentiviral vector carrying human LSR cDNA or vector only was used to infect HPDE6-C7 cells with LSR overexpression (LSR-OE) in cells. The efficiency of LSR-KD or LSR-OE was confirmed using RT-qPCR and Western blot. Moreover, to determine the effects of LSR on HPDE6-C7 cells in the HTG environment, cells were treated with 2 mM palmitic acid (PA) (Sigma, USA) for 24 h prior to use for subsequent experiments.