Figure 4.

Knockdown of SPP1 inhibits melanoma cell proliferation, migration and invasion. (A-B) SPP1 knockdown efficiency in A375 and SK-MEL-28 cell lines stably transduced with control shRNA (shCtrl) or two SPP1 shRNAs (shSPP1 #1 and #2) quantified by RT-PCR (A) and western blotting (B). (C-D) Cell proliferation of A375 (C) and SK-MEL-28 (D) cells transduced with shCtrl, shSPP1 #1, and shSPP1 #2 were quantified by CCK-8 assay. (E-F) Scratch-wound healing assay of A375 and SK-MEL-28 cells transduced with shCtrl, shSPP1 #1, and shSPP1 #2. Six random areas were selected. Images (at 40X magnification) were taken at 0, 24, and 48 hours. (G) Invasiveness of A375 cells transduced with shCtrl, shSPP1 #1, and shSPP1 #2 were assessed by Transwell assays. Invaded cells were determined for 18 hours. Five random areas were selected. Images were taken at 200X magnification. (H) model pattern and picture of resected subcutaneous xenografted tumors. A375 cells (10⁶) were injected subcutaneously in the right flank of NSG mice. Tumors were resected and photographed at day 21 (N = 6 in each group). (I) Tumor volume were recorded every other day and calculated as ([length×width²]/2). (J) Tumor weights were recorded after tumor resection at day 21. All data were represented as mean ± SD of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001.