Fig. 3

Silencing of CXCL5/CXCR2 axis inhibits migration and invasion in vitro and lung metastasis in vivo. a The relative mRNA levels of CXCL5/CXCR2 in the indicated stable cell lines were determined by qRT-PCR. The mean ± SD of triplicate samples are shown; n = 3. b The decreased expression of CXCL5 and CXCR2 in the CXCL5/CXCR2-silenced S18 and 5-8F cells compared with the scramble shRNA control cells were confirmed by western blotting. c The cell growth rates were compared between the CXCR2-, CXCL5-, CXCR2 + CXCL5- and scramble shRNA-transfected control cells using cell growth assays. d Representative images and summaries of the foci formation induced by cells transfected with CXCL5 and CXCR2 shRNA and scramble shRNA control cells. e The cell migration and invasion of the indicated stable cell lines were determined as described in the Materials and Methods. Representative images of the migrated and invaded cells are shown. f, g The indicated stable transfected cells were injected into the lateral tail veins of nude mice. The superior and inferior panels present the macroscopic appearance of metastatic lung tumours and H & E staining, respectively. The arrows indicate the metastatic nodules. Original magnification: 20 × objective. h, i The tumour weights and numbers of metastases per lung in the mice that were injected with the indicated stable cells, and the expression of human HPRT mRNA relative to mouse 18S rRNA in the lungs of the tumour-bearing nude mice are shown in H and I, respectively. There were 10 mice in each group. These experiments were repeated at least three times. Error bars, mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001