Figure 3
The effect of Flot-2 overexpression on the biological characteristics of 6-10B-Flot-2 cells.
A, MTT assay, colony formation assay and flow cytometric analysis were carried out to analyze the influence of ectopic Flot-2 expression on the growth, proliferation and cell cycle stage of 6-10B cells. Enhanced proliferation (significant differences were observed since Day3), colony formation (colony number: 6-10B-pcDNA3.1(+): 5.48 ± 1.44, 6-10B-Flot-2: 13.67 ± 2.45) and cell cycle progression(S stage ratio(%): 6-10B-pcDNA3.1(+): 36.01 ± 0.08, 6-10B-Flot-2: 20.54 ± 0.79) were observed for 6-10B-Flot-2 cells. B, Effects of Flot-2 on cell motility of 6-10B cells measured by an in vitro scratch wound healing assay. C, The influences of Flot-2 on the motility and in vitro invasiveness of 6-10B cells measured by a migration assay (cell number: 6-10B-pcDNA3.1 (+):16.7 ± 3.34, 6-10B-Flot-2: 46.58 ± 4.35) and an in vitro Matrigel invasion assay (cell number: 6-10B-pcDNA3.1 (+): 63.75 ± 8.13, 6-10B-Flot-2: 132.21 ± 17.63). 6-10B-Flot-2 cells displayed significantly stronger migratory and invasive capacities than that of 6-10B-pcDNA3.1 (+) cells in vitro. D, In vivo metastasis assay of 6-10B-Flot-2 cells. After intraperitoneal inoculation, 6-10B-Flot-2 cells invaded into diaphragmatic muscle, metastasized to lung and formed multiple metastases and metastasized to the mediastinal lymph nodes. Thus, the invasive and metastatic capacities of 6-10B-Flot-2 were also confirmed in vivo. All data were representative of three independent experiments. The data were analyzed by a two-tailed student t-test.* indicates P < 0.05.
