Figure 4.

miR-130a attenuates rapamycin and starvation-induced autophagy in ovarian cancer cells. (a, b) Autophagy-related markers (p-ULK1, LC3, Atg 7 and Beclin1 and p62) were tested by western blot in rapamycin/starvation treated A2780 cells with and without miR-130a overexpression. (c,d) Western blot analysis of LC3 expression in rapamycin or starvation treated A2780 cells transfected with miR-130a inhibitor or not. (e) Restoration of TSC1 could partly rescued miR-130a blocked autophagy in A2780 and SKOV3 cells treated with rapamycin (60 nM). (f) LC3-II and p62 expression was measured by western blot in A2780 cells with and without miR-130a overexpression in the presence or absence of lysosomal protease inhibitor E64D and PepA. (g) After transfection of A2780 cells with NC or miR-130a/anti-miR-130a, cells were infected with GFP-RFP-LC3 adenovirus for 24 h, then cells were dealt with starvation or rapamycin. Autophagic flux was monitored by confocal microscopy. The mean number of autophagosomes per cell (yellow puncta in merged images) and autolysosomes (red puncta in merged images) were counted. *P<0.05