Htert hpne

The normal pancreatic cell line hTERT-HPNE and two human PDAC cell lines (PANC-1, SW1990) were purchased from the American Type Culture Collection (ATCC) and the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). We cultured in RPMI 1640 (Thermo Fisher, Waltham, MA, USA) supplemented with 10% FBS (Thermo Fisher). hTERT-HPNE cells were maintained in DMEM medium supplemented with 5% FBS, human epidermal growth factor (EGF 10 ng/mL, ThermoFischer, Waltham, MA, USA), puromycin (750 ng/mL, ThermoFischer, Waltham, MA, USA) and 5 mM D-glucose. All cells were cultured in a humidified incubator containing 5% CO2 at 37 °C. Small interfering RNAs (siRNAs) targeting lncRNA DCST1-AS1 and negative control siRNA (si-NC) were synthesized by GenePharma. (Shanghai, China). Transfection was carried out using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) and the final concentration was 100 nm for siRNAs.

Human pancreatic cancer cell lines Capan-2, CFPAC-1, SW1990, Mia-PaCa-2, PATU-8988, PANC-1, and Normal pancreatic duct cell line hTERT-HPNE were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Capan-2 cells were cultured in McCoy’s 5 A medium (Thermo Fisher Scientific). CFPAC-1 cells were cultured in Iscove’s modified Dulbecco’s medium (IMDM, Thermo Fisher Scientific). SW1990, Mia-PaCa-2, PATU-8988, PANC-1, and hTERT-HPNE cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Thermo Fisher Scientific). All media were supplemented with 10% fetal bovine serum (Thermo Fisher Scientific), penicillin (100 U/mL), and streptomycin (100 µg/mL). Cells were incubated at 37 °C in an atmosphere of 5% CO₂. HEK-293 F cells were cultured in suspension with serum-free medium (Catalog No. 12338018; Thermo Fisher Scientific), and incubated at 37 °C in an atmosphere of 8% CO₂.

Human PC cell lines (PANC-1, CFPAC-1, BxPC-3, AsPC-1, MIA PaCa-2) and immortalized normal pancreatic duct epithelial hTERT-HPNE cells were obtained from the American Type Culture Collection (Manassas, VA, USA). Cells were cultured in Dulbecco’s modified Eagle’s medium (hTERT-HPNE, PANC-1, MIA PaCa-2), RPMI-1640 (CFPAC-1, BxPC-3), or IMDM (AsPC-1) medium supplemented with 10% fetal bovine serum (Thermo Scientific, Waltham, MA, USA) in a humidified atmosphere of 5% CO₂ at 37 °C. All cell lines were authenticated using standard short tandem repeat testing by Biowing Applied Biotechnology, Co., Ltd (Shanghai, China). For details about the generation of Gem-R cells, ABCA8-overexpressing cells, or ABCA8-knockdown cells, see Supplementary Materials and Methods.

Human pancreatic cancer cell lines, including AsPC-1, BxPC-3, MIA PaCa-2, PANC-1, T3M4 and normal pancreatic ductal cell (hTERT-HPNE) were purchased from the American Type Culture Collection (ATCC). Human pancreatic cancer cell line PaTu8988 was from DSMZ. Human umbilical vein endothelial cell (HUVEC) was purchased from the Pharmlab in china. The HEK-293T was generously provided by professor Mao in the department of Biochemistry and Molecular Biology in Peking university. All cell lines were authenticated by short tandem repeats (STR).
The hTERT-HPNE was cultured in in 75% DMEM without glucose (Gibco, USA) and 25% M3 Base (Incell, USA) supplemented with 10 ng/ml human recombinant EGF (CST, USA), 5.5 mM D-glucose (1 g/L), 5% fetal bovine serum (Gibco, USA), 0.75 mg/ml puromycin (sigma, USA). AsPC-1, BxPC-3, T3M4 were cultured in RPMI 1640 (Gibco, USA). MIA PaCa-2, PANC-1, PaTu8988, HEK-293T were cultured in DMEM. HUVEC was cultured in Endothelial Cell Medium (ScienCell, USA) supplemented with 5% fetal bovine serum, endothelial cell growth supplement, 1% penicillin-streptomycin solution (KeyGEN, china). Cells were maintained in cell incubator (ThermoFisher, USA) with 5% CO2 and 20% O2 for normoxic condition and incubator chamber (Billups-rothenberg, USA) flushed with 5% CO2 and 95% N2 for hypoxic condition.

The PANC-1, BXPC-3, CAPAN-2, MIAPACA-2, HPAC, CFPAC-1, PAN02, HAF, hTERT-HPNE, and 293T cell lines were purchased from ATCC and Chinese Academy of Sciences Cell Library. PANC-1, PAN02, MIAPACA-2, HAF and 293T cells were cultured in DMEM (Gibco). BXPC-3, CAPAN-2, and HPAC cells were cultured in RPMI-1640 (Gibco), CFPAC-1 cells were cultured in IMDM (Gibco). hTERT-HPNE cells were cultured in 75% DMEM lacking glucose and 25% medium M3 base and supplemented with 5% fetal bovine serum (FBS), 10 ng/ml human recombinant epidermal growth factor, 5.5 mM D-glucose, and 750 ng/ml puromycin. Except for hTERT-HPNE cells, the media used for the cells was supplemented with 10% FBS (Gibco) and 1% penicillin/streptomycin. The medium for MIAPACA-2 cells also contained 5% horse serum. All the cells were using standard culture conditions at 37 °C in an atmosphere of 5% CO₂.