Panc 1

Human pancreas ductal epithelioid (HPDE) cells and four human pancreatic cancer cell lines (SW1990, AsPC-1, BxPC-3, and PANC-1) were purchased from the American Type Culture Collection. Cells were grown in RPMI-1640 medium (Invitrogen, CA, USA) supplemented with 10% fetal bovine serum (Gibco, CA, USA) and cultured in a 37°C humidified atmosphere of 5% CO₂. The knockdown and overexpression of TUG1 in BxPC-3 and PANC-1 cells were achieved by transfection with lentivirus vector containing TUG1 shRNA (forward, 5′-GATCCGCTTGGCTTCTATTCTGAATCCTTTCAAGAGAAGGATTCAGAATAGAAAGCCAAGCCAAGCTTTTTTG-3′; reverse, 5′-GCGAACCGAAGATAAGACTTAGGAAAGTTCTCTTCCTAAGTCTTATCTTCGGTTCGAAAAAAC-3′; GenePharma, Shanghai, China), The overexpression of TUG1 in SW1990 cells was achieved by transfection with the TUG1-pcDNA3.1 plasmid which constructed by Invitrogen (Invitrogen, CA, USA). Cells were transfected by Lipofectamine 2000 (Invitrogen, CA, USA). The overexpression and knockdown of miR-29c were performed using miR-29c mimic and miR-29c inhibitor (GeneCopoeia, Guangzhou, China), respectively. Cells transfected with empty vector or scramble control were used as negative control. Cells were plated in 6-well clusters or 96-well plates and transfected for 24 or 48 h. Transfected cells were used for further assays or protein extraction.