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
Figure 2. CAFs secreted exo-miR-103a-3p to suppress apoptosis of NSCLC cells. (A) TEM images of exosomes isolated from NSCLC cell lines NCI-H1650 and NCI-H1299. (B) Western blot for CD63, Alix, and Tsg101 in exosomes from NCI-H1650 and NCI-H1299 cells. (C) Immunofluorescence staining of PKH-26 labelled CAF exosomes in NCI-H1650 and NCI-H1299 cells. DAPI indicates nucleus. Exosomes from NFs and CAFs transfected miR-103a-3p or AMO-miR-103a-3p or its NC were isolated, then NCI-H1650 and NCI-H1299 cells were incubated with NF or CAF exosomes. (D) qRT-PCR analyzed the expression of miR-103a-3p NCI-H1650 and NCI-H1299 cells. (E) CCK8 was used to test viability of NCI-H1650 and NCI-H1299 cells. (F, G) The expressions of apoptosis related protein Bax, Caspase3 and Caspase9 were analyzed by western bolt. (H, I) The number of apoptotic cells was calculated by flow cytometry. *p<0.05 vs NF exo or, CAF-miR-NC exo or CAF-AMO-miR-NC exo, # p<0.05 vs CAF-miR-NC exo or CAF-AMO-miR-NC exo.