fig5

Figure 5. MiR-204-5p/PFN2, miR-204-5p/BCL2 and miR-204-5p/SIRT1 axes were involved in exosomal LYPLAL1-DT mediates tumorigenesis and angiogenesis. (A and B) MiR-204-5p levels in H446 cells were measured by qPCR following treatment with exosomes from LYPLAL1-DT-overexpressing H446 cells (OE-Exo) (A) or HUVEC cells (OE-Exo) (B); (C and D) PFN2 mRNA expression was determined by qPCR in H446 (C) and DMS114 (D) cells after treatment with exosomes from LYPLAL1-DT-overexpressing-H446 cells (OE-Exo) or control cells (OC-Exo); (E) PFN2 expression was analyzed by qPCR in LYPLAL1-DT-knockdown H196 cells treated with exosomes from LYPLAL1-DT-overexpressing-H446 cells (OE-Exo) or control cells (OC-Exo); (F) PFN2 mRNA levels in H446 cells exposed to exosomes from LYPLAL1-DT-overexpressing HUVEC cells (OE-Exo) or control cells (OC-Exo); (G and H) PFN2 protein levels were detected by Western blotting in H446 cells treated with exosomes from LYPLAL1-DT-overexpressing H446 cells (OE-Exo) (G) or HUVEC cells (OE-Exo) (H); (I) The expression level of BCL2 was measured in H446 cells when co-cultured with LYPLAL1-DT-OC- or OE- H446 cells; (J-L) The expression level of BCL2 in H446, H196 and DMS114 cells when treated with exosomes from LYPLAL1-DT-overexpressing-H446 or control cells (OC-Exo); (M and N) The expression level of BCL2 in H446 cells treated with exosomes from LYPLAL1-DT-overexpressing-HUVEC cells (OE-Exo) or control cells (OC-Exo) was measured by qRT-PCR and Western blotting; (O-R) HUVEC cells treated with exosomes from LYPLAL1-DT-overexpressing-H446 cells (OE-Exo) or control cells (OC-Exo). The expression levels of miR-204-5p (O), SIRT1 (P and Q) and PFN2 (R) were tested by qPCR and Western blot. Data are presented as mean ± SD. Differences between the two groups were analyzed by Student’s t-test, while comparisons among more than two groups were analyzed by one-way ANOVA. (^*P < 0.05; ^**P < 0.01; ^***P < 0.001; ^****P < 0.0001). . MiR-204-5p: MicroRNA-204-5p; PFN2: profilin 2; BCL2: B-cell lymphoma 2; SIRT1: sirtuin 1; LYPLAL1-DT: LYPLAL1 divergent transcript; OE-Exo: exosomes from overexpressing cells; OC-Exo: exosomes from control cells; HUVEC: human umbilical vein endothelial cells; qPCR: quantitative polymerase chain reaction; SD: standard deviation; ANOVA: analysis of variance.