Research Paper Volume 11, Issue 15 pp 5705—5725

Figure 7. ADAMTS9-AS2 impedes ccRCC cell proliferation and chemoresistance by acting as a miR-27a-3p sponge. (A) MTS assays were performed in caki-1 cells transfected with NC or si-FOXO1 and treated with the indicated concentrations of 5-Fu or Cisplatin. (B) MTS assays were performed in caki-1 cells transfected with pcDNA control, pcDNA ADAMTS9-AS2, or pcDNA ADAMTS9-AS2 simultaneous with miR-27a-3p, respectively. (C) MTS assays were performed in caki-1 cells transfected with pcDNA control, pcDNA ADAMTS9-AS2 MUT or pcDNA ADAMTS9-AS2 MUT simultaneous with miR-27a-3p, respectively. (D) MTS assays were performed in caki-1 cells transfected with pcDNA control, pcDNA ADAMTS9-AS2, or pcDNA ADAMTS9-AS2 simultaneous with miR-27a-3p and treated with the indicated concentrations of 5-Fu or Cisplatin. (E) MTS assays were performed in caki-1 cells transfected with pcDNA control, pcDNA ADAMTS9-AS2 MUT, or pcDNA ADAMTS9-AS2 MUT simultaneous with miR-27a-3p and treated with the indicated concentrations of 5-Fu or Cisplatin. (F) The expression of FOXO1 at the protein level was determined in caki-1 cells transfected with pcDNA control, pcDNA ADAMTS9-AS2, or pcDNA ADAMTS9-AS2 simultaneous with miR-27a-3p by western blot analysis. (G) The expression of FOXO1 at protein level was determined in caki-1 cells transfected with pcDNA control, pcDNA ADAMTS9-AS2 MUT or pcDNA ADAMTS9-AS2 MUT simultaneous with miR-27a-3p by western blot analysis. Three independent experiments were performed and data shown are mean ± SD. Statistically significant differences are indicated as *, P<0.05, **, P<0.01, ***, P<0.001; ns, no significance; Student’s t-test among two groups; ANOVA among multiple groups. ADAMTS9-AS2, ADAM metallopeptidase with thrombospondin type 1 motif, 9 antisense RNA 2; ccRCC, clear cell renal cell carcinoma; miR-27a-3p, microRNA-27a-3p; MTS, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfop-henyl)-2H-tetrazoli-um); NC, negative control; si, small interfering; FOXO1, Forkhead Box Protein O1; MUT, mutant type; SD, standard deviation.