|
Fig. 4
LncRNA STEAP3-AS1 positively regulates STEAP3 to promote CRC progression. A-B Correlation analysis of relative RNA level of STEAP3 and STEAP3-AS1 in TCGA datasets (A) and CRC cell lines from CCLE (B), r = Pearson?s correlation coefficient. C Relative RNA level of STEAP3-AS1 and STEAP3 in control or STEAP3-AS1-knockdown DLD-1 (left) and SW480 (right) cells determined by qPCR. (sh#1, shSTEAP3-AS1 #1; sh#2, shSTEAP3-AS1 #2). D The expression level of STEAP3 protein in control or STEAP3-AS1-knockdown DLD-1 and SW480 cells was measured by western blotting. (sh#1, shSTEAP3-AS1 #1; sh#2, shSTEAP3-AS1 #2). E-F The protein levels of STEAP3 and HIF-1? (E), and relative RNA level of STEAP3 (F) in DLD-1 and SW480 cells under hypoxia at indicated time points. G qPCR analysis of relative RNA levels of STEAP3, HIF-1? and HIF-1? target genes in mCherry SW480-derived zebrafish xenograft models under normoxia or hypoxia. H-I Relative cell numbers at serial time points (H) and colony formation (I) of control and STEAP3-AS1-knockdown DLD-1 cells with or without replenishment of STEAP3. (sh#1, shSTEAP3-AS1 #1; sh#2, shSTEAP3-AS1 #2; OE, STEAP3-AS1 overexpression). J The efficiency of siRNA-mediated STEAP3 knockdown was measured by western blotting. K The colony formation of control and STEAP3-AS1-overexpressing DLD-1 and SW480 cells treated with or without STEAP3 siRNA. (OE, STEAP3-AS1 overexpression). L Wound healing assay (left) and relative migration distance (right) of control and STEAP3-AS1-knockdown DLD-1 cells with or without reintroduction of STEAP3. Scale bar: 100 ?m. (sh#1, shSTEAP3-AS1 #1; sh#2, shSTEAP3-AS1 #2). M Wound healing assay (left) and relative migration distance (right) of control and STEAP3-AS1-overexpressing DLD-1 cells treated with or without STEAP3 siRNA. Scale bar: 100 ?m. (OE, STEAP3-AS1 overexpression). N Migration and invasion of control and STEAP3-AS1-knockdown DLD-1 cells with or without reintroduction of STEAP3. Scale bar: 100 ?m. (sh#1, shSTEAP3-AS1 #1; sh#2, shSTEAP3-AS1 #2). Data are means ± s.d. and are representative of at least 3 independent experiments. (* P < 0.05, ** P < 0.01, and *** P < 0.001)