circPOLR1C Promotes the Development of Esophageal Cancer by Adsorbing miR-361-3p and Regulating Cancer Cell Apoptosis and Metastasis

Figure 3

circPOLR1C could act as a sponge for miR-361-3p and negatively regulated miR-361-3p, which was poorly expressed in EC tissues. (a) Whether circPOLR1C could bind to miRNA was tested by RNA immunoprecipitation (RIP). circANRIL was employed as a negative control. (b) The enrichment of circPOLR1C and its targeted miRNAs (top 6) was detected by the circRIP experiment. (c) The enrichment of circPOLR1C and miR-361-3p was analyzed by the biotin-labeled microRNA capture experiment. (d) The targeted binding sites of circPOLR1C and miR-361-3p in CircInteractome. (e, f) The dual-luciferase reporter assay was used to verify the targeting relationship between circPOLR1C and miR-361-3p. (g) Colocalization of circPOLR1C and miR-361-3p in cells was detected by the FISH experiment under 400 × magnification. (h, i) The effects of sicircPOLR1C and overexpressed circPOLR1C on miR-361-3p expression. U6 was utilized as the internal reference. (j) The expression of miR-361-3p in EC tissues and ANT was analyzed by qRT-PCR (n = 46). (k) Correlation analysis of circPOLR1C and miR-361-3p in EC. All experiments were independently performed in triplicate. ^### vs. AGO2; ^ΔΔΔ vs. biotin-miR-361-3p-WT; ^†† vs. IC; ^§§§ vs. mimic control (MC); vs. siNC; ^ ^ ^ vs. NC; ^ξξξ vs. ANT.

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