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. 2021 Feb 18;13:1673–1686. doi: 10.2147/CMAR.S295833

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© 2021 Wei et al.

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Circ_0006174 acted as a sponge of miR-138-5p. (A and B) Relative enrichments of circ_0006174, U6 and 18S rRNA in cytoplasm and nucleus were detected by RT-qPCR. (C and D) Ago2 RIP assay was employed to assess the combination of Ago2 and circ_0006174 in CRC cells. (E) The potential target miRNAs of circ_0006174 were predicted by Circbank and Starbase 2.0. (F and G) RNA pull down assay was performed to evaluate the target relationship between miRNAs and circ_0006174. (H) The online tool StarBase 2.0 was used to predict the binding sites between circ_0006174 and miR-138-5p. (I) RT-qPCR assay was performed to analyze miR-138-5p expression in CRC cells transfected with miR-138-5p or miR-NC. (J and K) Dual-luciferase reporter assay was performed to verify the interaction between miR-138-5p and circ_0006174. (L) Relative expression of miR-138-5p in CRC cells transfected with sh-circ #1 or sh-NC was detected by RT-qPCR. Experiments were performed in triplicate, and data were presented as means ± SD. *P<0.05.