Human PDAC cell lines PANC-1, AsPC-1, Capan-2 and SW1990 were purchased from the Cell Resource Center, Shanghai Institute of Biochemistry and Cell Biology at the Chinese Academy of Sciences and the normal control pancreatic ductal epithelial cell line hTERT-HPNE was obtained from American Type Culture Collection (ATCC). All cell lines were cultured as recommended in growth medium supplemented with 10% fetal bovine serum (FBS) and incubated at 37 °C with a humidified atmosphere of 5 % CO2.
Three small interfering RNAs (siRNAs) against human FAM83B were transfected into human PDAC cells using the DharmaFECT 1 siRNA transfection reagent (Thermo Scientific Dharmacon lnc. USA) according to the manufacturer's instruction, while nonspecific siRNA was applied as a negative control. Real-time PCR was performed to measure the knockdown efficiency of the three FAM83B-siRNAs, and the most efficient one was chosen for further experiment. All the experiments were conducted in the log phase of cell growth. All the siRNAs were designed and synthesized at Genepharm Technologies (Shanghai, China). The sequences targeting FAM83B were listed as follows:
5'- CUCGAGGAGUAUCUGUUUATT-3';
5'- GCCAUCUGAUAGUCUCAGUTT-3';
5'- GGGUCUCAGAAGUUAAGGUTT-3'.
Three small interfering RNAs (siRNAs) against human FAM83B were transfected into human PDAC cells using the DharmaFECT 1 siRNA transfection reagent (Thermo Scientific Dharmacon lnc. USA) according to the manufacturer's instruction, while nonspecific siRNA was applied as a negative control. Real-time PCR was performed to measure the knockdown efficiency of the three FAM83B-siRNAs, and the most efficient one was chosen for further experiment. All the experiments were conducted in the log phase of cell growth. All the siRNAs were designed and synthesized at Genepharm Technologies (Shanghai, China). The sequences targeting FAM83B were listed as follows:
5'- CUCGAGGAGUAUCUGUUUATT-3';
5'- GCCAUCUGAUAGUCUCAGUTT-3';
5'- GGGUCUCAGAAGUUAAGGUTT-3'.