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Fig. 7 | Molecular Cancer

Fig. 7

From: CircMET promotes tumor proliferation by enhancing CDKN2A mRNA decay and upregulating SMAD3

Fig. 7

CircMET functions as a ceRNA to sponge miRNAs. (A) Schematic of the selection for the direct downstream target of circMET. (B-C) The circMET-protein complex and miR-1197-Biotin complex were enriched by circMET junction probe or Biotin-antibody with protein extracts from UOK109 and 786-O cells. The enrichment of circMET and miR-1197 was detected by real-time PCR. (D) Model of AGO2-RIP assay. (E) RIP assays were performed using AGO2 antibody in UOK109 cells, then the enrichment of circMET was detected by real-time PCR. (F) Model of MS2-RIP assay (left). MS2-RIP-derived RNA was examined by real-time PCR (right). The levels of the real-time PCR products were normalized relative to IgG control. (G) Schematic illustration of circMET wild type and mutation luciferase reporter vectors. (H) HEK293T cells were co-transfected with miRNA mimics and wild-type or mutant circMET luciferase reporter vector, and luciferase reporter activity was detected. (I-J) The effect of circMET on miR-1197 expression in UOK109 cells was analyzed by real-time PCR after overexpression or knockdown of circMET. (K) Cell viability of UOK109 and 786-O cells was determined using CCK-8 assays after transfection for 48 h. (L-M) A colony formation and tumor sphere formation assay were used to determine the colony and tumor sphere formation ability of UOK109 and 786-O cells co-transfected with indicated lentivirus. (N) Cell cycle analysis was performed using flow cytometry in cells transfected with indicated lentivirus. The data are presented as the mean ± SD, n.s. = non-significant, **P < 0.01, ***P < 0.001

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