TPC1 cells and BCPAP cells were cultured in DMEM/F-12 medium (Gibco, Thermo Fisher Scientific) supplemented with 2 mM glutamine, 10% fetal bovine serum (Gibco, Thermo Fisher Scientific), and 100 U/ml penicillin/streptomycin (Sigma, St. Louis, MO, United States). The cells were maintained at 37°C in a humidified atmosphere containing 5% CO2. Rapamycin (Sirolimus) was purchased from Selleck (Shanghai, China).
The siRNAs of ENO1 and CST1 were purchased from GenePharma. siCtrl: 5′-UUCUCCGAACGUGUCACGU-3′; siENO1#1: 5′-CGUACCGCUUCCUUAGAACUU-3′; siENO1 #2:5′-GAAUGUCAUCAAGGAGAAAUA-3′; siCST1#1:5′-CC ACCAAAGAUGACUACUA-3′; siCST1#2:5′-GCUCUUUCGAG AUCUACGA-3′; siCST4#1:5′-CUUUCGAGAUCUACGAAGU UCTT-3′; siCST1#2:5′-CCAUCAGCGAGUACAACAATT-3′. According to the manufacturer’s protocol, Lipofectamine RNAiMAX transfection reagent (Invitrogen; Thermo Fisher Scientific) was used for transfection of siRNA. Lipofectamine 3000 (Thermo Fisher Scientific) was used for plasmid overexpression.
The siRNAs of ENO1 and CST1 were purchased from GenePharma. siCtrl: 5′-UUCUCCGAACGUGUCACGU-3′; siENO1#1: 5′-CGUACCGCUUCCUUAGAACUU-3′; siENO1 #2:5′-GAAUGUCAUCAAGGAGAAAUA-3′; siCST1#1:5′-CC ACCAAAGAUGACUACUA-3′; siCST1#2:5′-GCUCUUUCGAG AUCUACGA-3′; siCST4#1:5′-CUUUCGAGAUCUACGAAGU UCTT-3′; siCST1#2:5′-CCAUCAGCGAGUACAACAATT-3′. According to the manufacturer’s protocol, Lipofectamine RNAiMAX transfection reagent (Invitrogen; Thermo Fisher Scientific) was used for transfection of siRNA. Lipofectamine 3000 (Thermo Fisher Scientific) was used for plasmid overexpression.