Research Paper Volume 13, Issue 10 pp 14456—14468

Cancer-associated fibroblasts secreted miR-103a-3p suppresses apoptosis and promotes cisplatin resistance in non-small cell lung cancer

Pum2 mediated miR-103a-3p packaging into CAF exosomes. (A) RBPDB analysis of the specific interaction between miR-103a-3p and RBP motifs (threshold 0.8). Two siRNAs of Pum2 were transfected into CAFs, and the silencing efficiency was calculated by (B) western blot and (C) qRT-PCR assay. qRT-PCR assay was used to test the expression of miR-103a-3p in (D) CAFs and (E) exosomes from CAFs. (F) Western blot analysis of Pum2 expression in samples derived by miRNA pulldowns performed with nuclear, cytoplasmic, or exosomal CAFs lysates, biotinylated poly(G) was used as a negative control. (G) H1650 and NCI-H1299 cells were co-cultured with CAFs transfected with Cy3-miR-103a-3p and siRNA of Pum2 for 48 h. Fluorescence microscopy was used to detect red fluorescent signals. *p

Figure 3. Pum2 mediated miR-103a-3p packaging into CAF exosomes. (A) RBPDB analysis of the specific interaction between miR-103a-3p and RBP motifs (threshold 0.8). Two siRNAs of Pum2 were transfected into CAFs, and the silencing efficiency was calculated by (B) western blot and (C) qRT-PCR assay. qRT-PCR assay was used to test the expression of miR-103a-3p in (D) CAFs and (E) exosomes from CAFs. (F) Western blot analysis of Pum2 expression in samples derived by miRNA pulldowns performed with nuclear, cytoplasmic, or exosomal CAFs lysates, biotinylated poly(G) was used as a negative control. (G) H1650 and NCI-H1299 cells were co-cultured with CAFs transfected with Cy3-miR-103a-3p and siRNA of Pum2 for 48 h. Fluorescence microscopy was used to detect red fluorescent signals. *p<0.05 vs si-NC.