hsa_circ_0139402 Promotes Bladder Cancer Progression by Regulating hsa-miR-326/PAX8 Signaling

Figure 3

hsa_circ_0139402 acts as a sponge for hsa-miR-326, and PAX8 is a direct target of hsa-miR-326 in BC cell. (a) Bioinformatics analysis was used to search if hsa-miR-326 interacts with hsa_circ_0139402-MUT or hsa_circ_0139402-WT; (b) 4 miRNA mimics were cotransfected with the hsa_circ_0139402 vector into 5647 cells; the line means reduced at least half of the luciferase reporter activities; (c) RNA pull-down assay for the luciferase activity of hsa_circ_0139402-MUT or hsa_circ_0139402-WT in 5637 cells cotransfected with 6 miRNA mimics; (d) RIP assay for the amount of hsa_circ_0139402 and hsa-miR-326 in 5637 cells transfected with hsa_circ_0139402 overexpression or sh-hsa_circ_0139402 or negative control; (e) qRT-PCR analysis of expression levels of hsa-miR-326 in 5637 and UNUC3 cells transfected with hsa_circ_0139402 overexpression or sh-hsa_circ_01394021 or negative control; (f) immunofluorescence stain shows the interaction of hsa-mir-326 with hsa_circ_0139402; (g) luciferase reporter assay for the luciferase activity of PAX8-MUT or PAX8-WT in 5637 cells cotransfected with hsa-miR-326; (h) QRT-PCR analysis of the expression levels of hsa-miR-326 in BC tissues compared with normal tissues; (i) the expression levels of PAX8 in BC tumor tissues compared with adjacent normal tissues; (j) Pearson correlation was used for correlation analysis between hsa_circ_0139402, hsa-miR-326, and PAX8 in BC patients. Data were represented as . compared with negative control; ^# compared with hsa_circ_0139402+hsa-miR-326 mimics.

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