Extracellular Vesicle-Loaded Oncogenic lncRNA NEAT1 from Adipose-Derived Mesenchymal Stem Cells Confers Gemcitabine Resistance in Pancreatic Cancer via miR-491-5p/Snail/SOCS3 Axis

Figure 4

Verification of the binding between NEAT1 and miR-491-5p in PCa cells. (a) The binding site between NEAT1 and miR-491-5p in PCa cells predicted by the StarBase database. (b) The colocalization of NEAT1 and miR-491-5p detected by FISH (scale bar: 25 μm). (c) The relationship between miR-491-5p expression and the prognosis of PCa patients (44 patients with high miR-491-5p expression and 132 patients with low miR-491-5p expression) analyzed by Ualcan database. (d) miR-491-5p expression in dasatinib-sensitive PCa cell lines and dasatinib-resistant PCa cell lines analyzed in GSE59357 (). (e) Luciferase activity of NEAT1-WT/MUT analyzed by dual-luciferase reporter gene assay. (f) Enrichment of miR-491-5p by NEAT1 in SW1990 cells treated with anti-Ago2 or anti-IgG detected by RIP. (g) RNA pull-down assay for a biotinylated NEAT1 and an NC probe, miR-491-5p enrichment detected by RT-qPCR. (h) NEAT1 expression in SW1990 cells treated with overexpression vector or shRNA-mediated NEAT1 measured by RT-qPCR. (i) miR-491-5p expression in SW1990 cells treated with overexpression vector or shRNA-mediated NEAT1 measured by RT-qPCR. (j) miR-491-5p expression in SW1990 cells cocultured with EVs from sh-NEAT1-treated ADSCs measured by RT-qPCR. vs. SW1990 cells transduced with mimic NC or inhibitor NC, SW1990 cells treated with Bio-NC-probe, Vector, or PBS; ^# vs. SW1990 cells transduced with sh-NC or SW1990 cells cocultured with EVs from ADSCs transduced with sh-NC. All data were presented as deviation. Comparisons of data between two groups were analyzed by unpaired -test, and comparison of data among multiple groups was tested by one-way analysis of variance, followed by Tukey’s post hoc test. Cell experiments were independently repeated three times.

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