Circular RNA-ITCH Suppresses Lung Cancer Proliferation via Inhibiting the Wnt/β-Catenin Pathway

Figure 4

cir-ITCH can regulate the Wnt/β-catenin signaling pathway and cellular proliferation of lung cancer cells. (a) A549 and NCI-H460 cells were transfected with cir-ITCH or empty control vector; then, a β-catenin/T-cell factor- (TCF-) responsive luciferase reporter assay was performed. The luciferase activity was normalized to the Renilla luciferase activity. (b) The protein levels of Wnt3a and β-catenin were assessed in A549 cells and NCI-H460 cells by western blotting. β-actin was used as endogenous control. (c) The mRNA level of c-Myc and cyclinD1 in A549 cells and NCI-H460 cells was detected by qRT-PCR after being transfected with cir-ITCH or empty control vector. (d) A549 and (e) NCI-H460 cells were seeded in 96-well plates after being transfected with cir-ITCH and empty control vector, and cell proliferation was detected daily for 3 days by using the CCK-8 assay. Data are presented as mean ± SEM from six replicates for each group. .

(a)

(b)

(c)

(d)

(e)