Fig. 5

miR-17-5p inhibits autophagy and sunitinib resistance in RC cells. miR-17-5p gain and loss of function was performed in RC cells, in which parental cells were transfected with miR-17-5p inhibitor, and sunitinib-resistant cells were transfected with miR-17-5p mimic. After cell transfection, the mRNA expression of miR-17-5p was assayed by qRT-PCR (a). The mRNA and protein expressions of Beclin1 (b, c), LC3-II/LC3-I (f), and p62 (f) were tested by Western blot and qRT-PCR. Then miR-17-5p was found to negatively mediate Beclin1. MDC staining observed more formation of autophagic vacuoles in parental cells transfected with miR-17-5p inhibitor. Overexpression of miR-17-5p lessened the number of autophagic vacuole formation of sunitinib-resistant cells (d). After transfection, immunofluorescence revealed increased LC3 in parental cells and diminished expression intensity of LC3 in sunitinib-resistant cells (e). CCK-8 assay detection on the sensitivity of RC cells to sunitinib unfolded that parental cells transfected with miR-17-5p inhibitor had improved sunitinib resistance, while sunitinib-resistant cells transfected with miR-17-5p mimic possessed reduced sunitinib resistance of RC cells (g). The colony formation on cell colony formation ability exhibited that knockdown of miR-17-5p in 786-O or ACHN cells elevated clone number, whereas overexpression of miR-17-5p in 786-O-R or ACHN-R cells declined clone number (h); ^*P < 0.05, ^**P < 0.01, compared to 786-O + inhibitor NC or 786-O-R + mimic NC group; ^#P < 0.05, ^##P < 0.01, compared to ACHN + inhibitor NC or ACHN-R + mimic NC group; RC, renal cancer