Figure 1.

OPN is highly expressed by NSCLC cells and promotes NSCLC cell EMT.

Notes: (A) The mRNA expression levels of OPN in NSCLC cell lines (A549, NCI-H358, H1299, H460) and normal lung epithelium cell line (BEAS-2B) were examined by real-time qPCR. GAPDH was used as an internal control. The mRNA expression of OPN measured in BEAS-2B is presented as 1. (B) The protein secretion levels of OPN were examined by ELISA. The detection range of the kit is 0.3–90 ng/mL. (C–E) Real-time qPCR was used to examine the mRNA levels of E-cadherin, N-cadherin and Vimentin in A549 and NCI-H358 cells treated with an increasing concentration of OPN (0, 100, 200 and 400 ng/mL) for 24 hours. GAPDH was used as an internal control. The mRNA expression of target genes in control group is presented as 1. Results presented are mean ± SD analyzed with Student’s t-test from three independent experiments. *P<0.05, **P<0.01, ***P<0.001 versus control. (F and G) Western blot was used to detect the protein levels of E-cadherin, N-cadherin and Vimentin in A549 and NCI-H358 cells treated with an increasing concentration of OPN (0, 100, 200 and 400 ng/mL) for 24 hours. β-Actin was used as an internal control. The group treated with control buffer served as control.

Abbreviations: OPN, osteopontin; NSCLC, non-small cell lung cancer; EMT, epithelial-mesenchymal transition; ELISA, enzyme linked immunosorbent assay; qPCR, quantitative polymerase chain reaction; SD, standard deviation.