LINC00888 directly suppressed miR-126/CRK axis in A375-S2. (A) The predicted binding sites between LINC00888 and miR-126-5p through complementary base-pairs. The wild-type LINC00888 3′UTR (WT; containing the binding sites of miR-126-5p) or mutant-type LINC00888 3′UTR (MT) vectors were constructed. (B and C) A375-S2 cells were treated with LINC00888 siRNA2 or lenti-LINC00888 for 72 hours, and the expression of miR-126 were detected by qRT-PCR (n=3, **p<0.01 vs control). (D) miR-126 could directly regulate transcriptional level of LINC00888, demonstrated by dual-luciferase reporter assay (n=3, **p<0.01 vs control). (E) The predicted binding sites between targeted gene CRK and miR-126-5p through complementary base-pairs. The wild-type CRK 3′UTR (WT) or mutant-type CRK 3′UTR (MT) vectors containing the binding sites of miR-126-5p were constructed. (F) LINC00888 stable knockdown and control A375-S2 cells were cotransfected with miR-126 inhibitors and luciferase reporters containing CRK 3′UTR, or nothing. Luciferase activities were then detected by dual-luciferase reporter assay (n=3, **p<0.01 vs control). (G) LINC00888 stable overexpression and control A375-S2 cells were cotransfected with miR-126 inhibitors and luciferase reporters containing CRK 3′UTR, or nothing. Luciferase activities were then detected by dual-luciferase reporter assay (n=3, **p<0.01 vs control).
Abbreviations: lenti-LINC0088, lentivirus LINC00888; miR-126, microRNA-126; qRT-PCR, quantitative real-time polymerase chain reaction; UTR, untranslated region.