Fig. 4

HOTAIR regulates the expression of Beclin1 via binding to miR-17-5p. The binding sites of miR-17-5p and HOTAIR or miR-17-5p and Beclin1 were predicted. Then the wild type sequences of HOTAIR and Beclin1, as well as the mutated type sequences of HOTAIR and Beclin1 were accordingly designed (a). After sh-HOTAIR was transfected into 786-O-R, or ACHN-R cells, the expressions of miR-17-5p and Beclin1 were assessed by qRT-PCR and Western blot. The expression of miR-17-5p in sunitinib-resistant cells was lower than that in parental cells. Transfection with sh-HOTAIR enhanced the mRNA expression of miR-17-5p (b). The sunitinib-resistant cells had higher mRNA and protein levels of Beclin1 than parental cells. Knockdown of HOTAIR suppressed Beclin1 (c, d). The dual-luciferase reporter gene and RIP assay were employed to inspect the binding sites of HOTAIR and miR-17-5p as well as miR-17-5p and Beclin1 (e–g). ^*P < 0.05, ^**P < 0.01, compared to 786-O or ACHN or mimic NC or IgG group; ^#P < 0.05, ^##P < 0.01, compared to 786-O-R or ACHN-R group; RIP, RNA immunoprecipitation