Figure 4. CREB1, CCAR1 and JNK1 were the direct targets of miR-433. (a) Bioinformatics analyses showed overexpression of CREB1 (i), CCAR1 (ii) and JNK1 (iii) in the Oncomine database. (b) Real-time PCR manifested upregulation of CREB1 (i), CCAR1 (ii) and JNK1 (iii) in 35 paired CRC and normal mucosa specimens. (c) Bi-variable correlation analyses revealed that CREB1 (i), CCAR1 (ii) and JNK1 (iii) were all significantly correlated with miR-433 in 35 paired tissues. (d) A schematic of the putative binding sites between miR-433 and its targets according to the TargetScan database. (e) Western blot illustrating that miR-433 inhibited the expression of CREB1, CCAR1 and JNK1 in LoVo (i) and RKO (ii) cells. (f) Dual luciferase assays showing that miR-433 directly targets CREB1 (i), CCAR1 (ii) and JNK1 (iii). (g) Kaplan-Meier analysis combined with the log-rank method indicated that CRC patients with higher expression of CREB1 (i), CCAR1(ii) or JNK1 (iii) had a poorer prognosis. *, p<0.05; **, p<0.01; ***, p<0.001.