Fig. 4

Overexpression of miR-509-3p inhibited stemness, cisplatin resistance and cell growth, and promoted apoptosis of A549-dereived stem cells. A qRT-PCR analysis on miR-509-3pin spheres of A549-derived stem cells, *p < 0.01 compared with parental group. B qRT-PCR on the expression of stemness-associated gens (SOX2, CD44, and CD133) in A549cell spheres 48 h after transfection with mimic miRNAs, *p < 0.01 compared with Mock. C, D Number of tumor spheres in miR-509-3p overexpressing A459 stem cells, *p < 0.01 compared with Mock. E Flow cytometry analysis of cell cycle in A549-dereived stem cells 48 h after overexpression of miR-509-3p, *p < 0.01 compared with Mock. F qRT-PCR analysis of miR-509-3pin A549 and cisplatin-resistant A549/R cells. *p < 0.01 compared with A549. G CCK-8 assay on the cell viability of miR-509-3p-overexpressing A549-dereived stem cells treated with cisplatin at various concentration (0 μM, 4 μM, 8 μM, 16 μM, 32 μM), *p < 0.01 compared with Mock. H, I Annexin V-FITC/PI staining of A549-dereived stem cells 48 h after miR-509-3p overexpression, *p < 0.01 compared with Mock. J Western blotting analysis on expression of apoptosis-associated genes (Bax, Bcl-2 and cleaved-caspase-3) 48 h after transfection with miR-509-3p or Mock, *p < 0.01 compared with Mock. K, L Colony formation assay on cell growth of A549-dereived stem cells after 15 days culture, *p < 0.01 compared with Mock. M qRT-PCR analysis on proliferation-associated genes (Cyclin A1, Cyclin B1, and PCNA) 48 h after overexpression of miR-509-3p in A549-dereived stem cells, *p < 0.01 compared with Mock