LncRNA CAR10 Upregulates PDPK1 to Promote Cervical Cancer Development by Sponging miR-125b-5p

Figure 5

CAR10 acted as a ceRNA and upregulated the expression of PDPK1 by sponging miR-125b-5p. (a) The binding site of miR-125b-5p to the 3′UTR of PDPK1 was predicted by bioinformatics, and the binding site was mutated to construct luciferase reporter gene plasmids. (b, c) The effect of anti-miR-125b-5p on the activity of PDPK1 3′UTR WT and PDPK1 3′ UTR Mut was examined by luciferase assay in C33A and HeLa cells, , compared with Mock. (d, e, f) The effects of anti-miR-125b-5p on the expression of PDPK1 mRNA and protein were detected by RT-qPCR and western blot, , compared with Mock. (g) The mRNA expression level of PDPK1 was detected by RT-qPCR in the collected cervical cancer tissues and adjacent tissues, , compared with adjacent controls. The miR-125b-5p mimic (miR-125b-5p) and the negative control Mock were synthesized and transfected into C33A and HeLa cells, and (h) the expression level of miR-125b-5p was detected by RT-qPCR, , compared with Mock. (i, j) The effect of miR-125b-5p and/or CAR10 overexpression on the activity of PDPK1 3′UTR WT and PDPK1 3′ UTR Mut was examined by luciferase assay, , compared with CON. ^Ф, compared with CAR10 + miR-125b-5p. (k) The effect of miR-125b-5p and/or CAR10 overexpression on PDPK1 expression was observed by western blot and RT-qPCR, , compared with CON. ^Ф, compared with CAR10 + miR-125b-5p.

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