To further confirm the role of CsGRN in human cancer progression, wound-healing assays were performed to evaluate the effect of CsGRN on cell migration as described previously . The transfected PLC and RBE cells seeded in 6-well plates were grown to 80% confluence and were wounded by scratching with p200 pipette tips. Wounds were observed and photographed under a light microscope (Leica DMI3000B, Wetzlar, Germany) every 24 h for 72 h. The distances between the parallel cell edges were measured at each time point using Image J software. For each well, three different fields along the scratch were analysed in triplicate. Cell motility was measured as the percentage of the cell migration distance, which was regarded as the initial scratch distance. To evaluate the effect of CsGRN on cell invasion, we performed transwell assays as described previously . The PLC-GFP/RBE-GFP cells and PLC-GRN/RBE-GRN cells were suspended at 5.0 × 104 per insert with serum-free media and then were transferred to wells filled with a culture medium containing 10% FBS as a chemoattractant. After 24 h of incubation, non-invading cells on the top of the membrane were removed with a cotton swab. The migrated cells on the underside of the filter membrane were fixed and stained with 0.1% crystal violet. The number of migrated cells on the membrane was counted in five randomly selected microscopic fields, and the cells were photographed. The protocol used for the invasion assay was the same as that used for the migration assay, except that the transwell insert was coated with Matrigel (BD Biosciences, Heidelberg, Germany).