Fig. 5

Overexpression of ASAP1-IT1 blocked miR-509-3p-mediated cell functions in cancer stem cells both in vitro and in vivo. A Number of A549 cell tumor spheres after expression of Mock, pcDNA, miR-509-3p, and ASAP1-IT1, *p < 0.01 compared with Mock + pcDNA, φp < 0.01 compared with miR-509-3p + pcDNA, Фp < 0.01 compared with miR-509-3p + ASAP1-IT1. B qRT-PCR analysis on stemness-related genes (SOX2, CD44, and CD133) in A549 stem cells, *p < 0.01 compared with Mock + pcDNA, φp < 0.01compared with miR-509-3p + pcDNA. C Annexin V-FITC/PI staining of A549 stem cells with Mock, pcDNA, miR-509-3p, and ASAP1-IT1, *p < 0.01 compared with Mock + pcDNA, φp < 0.01 compared with miR-509-3p + pcDNA. D Western blotting analysis on apoptosis-associated genes (Bax, Bcl-2, and cleaved-caspase-3) in A549 stem cells, *p < 0.01 compared with Mock + pcDNA, φp < 0.01 compared with miR-509-3p + pcDNA. E Colony formation assay on cell growth after 15 days culture, *p < 0.01 compared with Mock + pcDNA, φp < 0.01 compared with miR-509-3p + pcDNA. F qRT-PCR analysis of growth-associated genes (Cyclin A1, Cyclin B1, and PCNA) in A549 stem cells, *p < 0.01 compared with Mock + pcDNA, φp < 0.01 compared with miR-509-3p + pcDNA. G Tumor volume in nude mice at the indicated time after inoculation, *p < 0.01 compared with Mock + pcDNA, φp < 0.01 compared with miR-509-3p + pcDNA. H Photo of harvested tumors at the 24th day after inoculation. I Tumor weight at the 24th day after inoculation, *p < 0.01 compared with Mock + pcDNA, φp < 0.01 compared with miR-509-3p + pcDNA. J qRT-PCR analysis on stemness-associated genes (SOX2, CD44, and CD133), apoptosis-associated genes (Bax and Bcl-2), and growth-associated genes (Cyclin A1, Cyclin B1, and PCNA) in tumors, *p < 0.01 compared with Mock + pcDNA, φp < 0.01 compared with miR-509-3p + pcDNA