Figure 3.

MiR-221 suppression-induced inhibition of invasion and migration is autophagy dependent. (A) Relative expression of miR-221 after overexpression and knockdown of miR-221 in 5637 and T24 cells detected by RT-qPCR. (B) Validation of the expression of TP53INP1, p-ERK1/2, and ERK1/2 using Western blot analysis. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was detected as loading control. (C) T24 cells were transfected with Lenti-sh-miR-221 or siRNA targeting ATG5 (siATG5). (D,E) Transwell invasion assays and wound healing assay of 5637 cells treated with Lenti-miR-221 and ERK inhibitor U0126 and T24 cells treated with Lenti-sh-miR-221 or siRNA targeting ATG5. Scale bar, 100 μm. (F) For the in vivo analyses, 5 × 10⁶ T24-sh-miR-221 cells were injected subcutaneously into the posterior hip of nude mice. The mice were continuously observed for 30 days. Images of the tumors generated by T24-sh-miR-221 cells (n = 4). Tumor volume was monitored every week. Tumor weight was evaluated in T24-sh-miR-221 or miR-NC treated mice. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001.