Figure 1.
DMH1 inhibits basal Smad phosphorylation and transcription of Id genes in NSCLC cells.
Chemical structure of DMH1; (B) Western blotting for phosphorylated Smad1/5/8 in A549 cells treated with DMH1 at various concentrations (1, 3, and 5 µM); (C) QPCR results of Id1, 2 and 3 in A549 cells treated with DMH1 and vehicle DMSO. *: p-value is <0.05.
Figure 2.
DMH1 decreases NSCLC cell migration and invasion.
Representative images from scratch-wound assays performed in the cultured A549 cells treated with DMSO, BMP4 and DMH1 (1 µM and 3 µM) for 24 hours. (B) Cell migrations of A549 cells were quantified by the gap distances after 24 hour treatment normalized with the initial gap distances. (C) Cell migrations of H460 cells were quantified in a similar way. (D) The effect of DMH1 (3 µM) on A549 cell invasion was determined using modified Boyden chamber assay in a 24-Multiwell Insert System (8 µM membrane, BD Biosciences) coated with matrigel. *: p-value is <0.05.
Figure 3.
DMH1 reduces A549 cell proliferation and induces cell death.
A549 cells were treated with DMH1 (3 and 5 µM) or DMSO for 48 hours and cell proliferation was determined by the sulforhodamine B (SRB) assay. (B) A549 cells were treated with DMH1 or DMSO for 72 hours, and cells were harvested for cell death assay using Trypan Blue staining. *: p-value is <0.05.
Figure 4.
DMH1 attenuates xenograft lung tumor growth in mice.
The A549 cells were subcutaneously implanted into the SCID mice followed by intraperitoneal injection of the vehicle (12.5% 2-hydroxypropyl-β-cyclodextrin) or 5 mg/kg DMH1 every other day for 4 weeks. Tumor volumes were measured from the sixth day to four weeks after injection. (B) Representative tumor tissues from mice treated with the vehicle control and DMH1 are compared. (C) Tumor tissues were stained for human specific Ki67 proliferation marker. *: p-value is <0.05.