Fig. 5.

circPTPRA suppresses EMT in NSCLC cell lines by sequestering miR-96-5p.

(a and c) circPTPRA-KD H23 cells and circPTPRA-KD H1755 cells exhibit heightened (a) migration and (c) invasiveness in a scratch and Transwell assay, respectively; co-transfection with miR-96-5p inhibitor reversed the phenotype; scale bar = 100 μm. *p < .05, **p < .01 vs. shCtrl, †p < .05, ††p < .01 vs. sh-circPTPRA [unpaired Student's t-test]. (b and d) circPTPRA-OE H522 cells and circPTPRA-OE H1755 cells exhibit diminished (b) migration and (d) invasiveness in a scratch and Transwell assay, respectively; co-transfection with miR-96-5p mimics reversed the phenotype; scale bar = 100 μm. *p < .05, **p < .01 vs. p-Ctrl, ††p < .01 vs. p-circPTPRA [unpaired Student's t-test]. (e and f) In vivo xenografts into nude BALB/c mice generated by tail-vein injection of stably-transfected H23 cells were sacrificed 6 weeks later to assess lung lesions (n = 9 mice per group). (e) Injection of circPTPRA-KD H23 cells promotes metastasis, which is reversed by co-transfection of miR-96-5p inhibitor, while (f) injection of circPTPRA-OE H23 cells suppresses metastasis, which is reversed by co-transfection of miR-96-5p mimics; *p < .05, **p < .01 vs. p-Ctrl, †p < .05, ††p < .01 vs. sh-circPTPRA or p-circPTPRA [unpaired Student's t-test]. Left panels: Typical lung images with metastatic lesions; middle panels: H&E lung tissue sections of metastatic lesions (100× magnification; scale bar = 100 μm); lower right panel: Quantification of metastatic lesions.

All in vitro experiments: n = 3 biological replicates × 3 technical replicates. Data presented as means with error bars representing standard errors of the mean (SEMs). Abbreviations: circPTPRA-KD = NSCLC cells with shRNA-mediated circPTPRA knockdown, circPTPRA-OE = NSCLC cells with circPTPRA overexpression plasmid, H&E = hematoxylin and eosin, ICC = immunocytochemistry, WB = Western blot.