Figure 2.

Pellino-1 overexpression enhances the cellular proliferation, migration, invasion and oncogenic transformation in A549 cells. (a) A549 cells were transfected with Myc (control), Myc-tagged Pellino-1, shLuc (control), Pellino-1-targeted shRNA (shPellino-1) or 3′-UTR Pellino-1-targeted shRNA (3′-UTR shPellino-1), and subjected to immunoblotting for Pellino-1 and actin. (b) MTT assay was performed to estimate the cell proliferation of A549 cells with Pellino-1 overexpression or knock-down. (c and d) A549 cells with Pellino-1 overexpression (Myc-Pellino-1) or Pellino-1 depletion (shPellino-1 and 3′-UTR shPellino-1) were pretreated with 10 μg/ml mitomycin C for 1 h at 37 °C, washed twice with PBS and then were subjected to transwell migration (c) and invasion assays (d). (e) A549 cells with Pellino-1 overexpression or Pellino-1 depletion were subjected to colony-forming assay. The numbers of colonies were counted in four randomly selected microscopic fields per plates. (f-g) A549 cells (5 × 10⁶) transfected with shLuc, shPellino-1 or 3′-UTR shPellino-1 were inoculated subcutaneously into the flank of athymic nude mice (five mice per group). Animals were monitored up to 33 days and tumor size was measured using an electronic caliper at 3-day intervals. All data are shown as the means±S.D. of at least three independent experiments. The P-values were calculated using unpaired Student's t-test. *P <0.05; **P <0.01; ***P <0.005