Figure 2.

Inhibition of MTBP decreases invasion and migration ability of lung adenocarcinoma cells both in vitro and in vivo.

Notes: (A) The expression level of MTBP protein measured in lung adenocarcinoma cell lines by Western blot. (B) siRNA1 inhibition of the MTBP expression in A549 cell line. (C and D) The expression level of MTBP mRNA and protein after shRNA lentiviruses treatment of A549 and H1975 cells analyzed by qPCR and Western blot. (E) Representative images of Transwell migration assays (top panel) and Transwell invasion assays (bottom panel) in A549 and H1975 cells. (F) Average number of migrated or invasive cell numbers per field of view ± SD from three independent experiments. (G) Representative images of cell migration analyzed by wound healing assay in A549 and H1975 cells. Bar: 100 µm. (H) Migratory distance of cells that migrated into the scratch wound at 6, 24, and 48 hours, calculated by the formula: Migration distance (µm)= width at 0 hour – width at observation point. (I and J) Representative image of tumors obtained from CAM inoculated with A549 and H1975 cells and tumor weights. (K and L) Representative images of metastatic cells in the lungs of chicken embryos and the number of metastatic cells in the lungs of chicken embryos observed with a confocal laser scanning microscope. Data in (C, F and H) are the mean ± SD of results from three independent experiments. Data in (J and L) are the mean ± SD of results for each group (Student’s t-test, *P<0.05, **P<0.01, and ***P<0.001).

Abbreviations: CAM, chorioallantoic membrane; MTBP, MDM₂ binding protein; NC, normal control; qPCR, quantitative PCR; WT, wild type.