Fig. 5
STXBP5-AS1 inhibited stem cell-like properties of PC cells by suppressing ADGB expression. a, b The expression level of ADGB in PANC-1 and Mia PaCa-2 cells co-transfected with two empty pSin vector (pSin-vector + pSin-Ctr), STXBP5-AS1 overexpression plasmid and an empty pSin vector (pSin-STXBP5-AS1 + pSin-Ctr) or STXBP5-AS1 overexpression plasmid and ADGB overexpression plasmid (pSin-STXBP5-AS1 + pSin-ADGB) was determined by qRT-PCR and western blot. c, d The expression level of ADGB in PANC-1/GR and Mia PaCa-2/GR cells co-transfected with two empty pSin vector (pSin-vector + pSin-Ctr), STXBP5-AS1 overexpression plasmid and an empty pSin vector (pSin-STXBP5-AS1 + pSin-Ctr) or STXBP5-AS1 overexpression plasmid and ADGB overexpression plasmid (pSin-STXBP5-AS1 + pSin-ADGB) was determined by qRT-PCR and western blot. e MTT assay showed that overexpressing ADGB rescued the decreased Gemcitabine IC50 in PANC-1/GR and Mia PaCa-2/GR cells due to forced expression of STXBP5-AS1. f Colony formation assay showed overexpressing ADGB rescued the decreased cell survival of Gemcitabine-challenged (50 ng/ml) PANC-1/GR and Mia PaCa-2/GR cells due to forced expression of STXBP5-AS1. g, h Caspase-3 activity assay and FACS indicated that overexpressing ADGB rescued the increased apoptosis of Gemcitabine-challenged (50 ng/ml) PANC-1/GR and Mia PaCa-2/GR cells due to forced expression of STXBP5-AS1. i Transwell assay showed overexpressing ADGB rescued the decreased cell invasion of PANC-1 and Mia PaCa-2 cells due to forced expression of STXBP5-AS1. j Tumor sphere formation assay showed overexpressing ADGB rescued the decreased spheres of PANC-1 and Mia PaCa-2 cells due to forced expression of STXBP5-AS1. k–m qRT-PCR and western blot indicated that overexpressing ADGB rescued the decreased expression levels of stem markers (Sox2, Bmi1, Lin28 and Nanog) in PANC-1 and Mia PaCa-2 cells due to forced expression of STXBP5-AS1. The data represent the mean ± SD from three independent experiments. *P < 0.05; ** P < 0.01, Student’s t test