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. 2021 Jul 12;13:5573–5585. doi: 10.2147/CMAR.S307595

Figure 5.

Figure 5

CircHIPK3 mediated HK2 expression through miR-1286. The levels of HK2 mRNA and protein by qRT-PCR and Western blot in BC tissues and adjacent normal breast tissues (A and B), BC tissues from primary patients (PTX-sensitive tissues) and recurrent patients (PTX-resistant tissues) after treatment based on PTX chemotherapy (C and D), MCF-10A, MDA-MB-231, MCF-7, MDA-MB-231/PTX and MCF-7/PTX cell lines (E and F). (G and H) Correlation between HK2 mRNA level and circHIPK3 or miR-1286 expression in BC tissues from 25 recurrent patients (PTX-resistant tissues) after treatment based on PTX chemotherapy. (I) Sequence of miR-1286, the potential miR-1286-binding sequence within HK2 3ʹUTR and the mutation of the seed region. (J and K) Dual-luciferase reporter assays in MDA-MB-231/PTX and MCF-7/PTX cells using HK2 3ʹUTR wild-type (HK2 3ʹUTR WT) or mutant (HK2 3ʹUTR MUT) luciferase reporters. (L–N) RIP experiments in MDA-MB-231/PTX and MCF-7/PTX cells using anti-Ago2 or anti-IgG antibody. (O–Q) HK2 protein level by Western blot in cells transfected with si-NC, si-circHIPK3#3, si-circHIPK3#3+in-miR-NC or si-circHIPK3#3+in-miR-1286. (A) adjacent normal breast tissues, T: tumor tissues, S: PTX-sensitive tumor tissues, R: PTX-resistant tumor tissues. *P < 0.05, **P < 0.01 or ***P < 0.001 by the Mann–Whitney U-test, Student’s t-test or ANOVA with Tukey’s post hoc test.