The Antitumor Effect of Gekko Sulfated Glycopeptide by Inhibiting bFGF-Induced Lymphangiogenesis

Figure 2

GSPP inhibits bFGF-induced migration of hLECs. (a) Wound healing assay. Left, representative images of injury width in wound healing assay (×40). Right, quantification of the migration ratio of hLECs compared to control. hLECs (1 × 10⁵ cells/well) were seeded in 24-well plates and wounds were generated after cell confluence. After hLECs were treated with different concentrations of GSPP (0, 10, and 100 μg/mL) and/or bFGF (10 ng/mL) for 0 h and 6 h, the photos were taken and the injury width was measured. The migration ratio was calculated as the migration width of experiment group/the migration width of control group. (b) Transwell assays. Left, representative images of migrated LECs in transwell assay (×40). Right, quantification of migrated LECs compared to control. hLECs (5 × 10⁴ cells/well) in EBM-2 with different concentrations of GSPP (0, 10, and 100 μg/mL) were added to the upper chamber of the transwell insert. EBM-2 containing bFGF (10 ng/mL) or not was added to the lower chamber to induce cell migration. After 12 h at 37°C, cells on the top surface of the membranes were wiped off with cotton balls, and the cells that migrated on the underside of inserts were fixed with methanol and stained with crystal violet. Five different digital images were taken per well, and the number of migrated cells was counted. (A) Control, (B) bFGF 10 ng/mL, (C) GSPP 10 µg/mL, (D) GSPP 100 µg/mL, (E) GSPP 10 µg/mL + bFGF 10 ng/mL, and (F) GSPP 100 µg/mL + bFGF 10 ng/mL. Data were presented as mean ± SD of three independent experiments; versus control group and versus bFGF-single use group.

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