Figure 2. Sema3A promoted proliferation, migration, and invasion of HCC cells in vitro and promoted gene changes between stably transfected and parent cells.

A. Sema3A expression was confirmed using qRT-PCR analysis of stably transfected and of parent cells. B. ELISA results indicated the presence of significantly greater levels of Sema3A proteins in PLC/PRF/5-Sema3A and HCCLM3-Mock cells, compared with PLC/PRF/5-Mock and HCCLM3-shRNA-Sema3A. The mean (± SD) values from three independent experiments are presented, **p<0.01. C. CCK8 assay was used to detect cell proliferation, *p<0.05; **p<0.01. D. The results for wound-healing migration assays and the relative quantification of percent open area are presented. Results shown are mean (±SD) values from three independent experiments, **p<0.01. Magnification, ×200. E. Transwell Matrigel invasion assays were used to test invasive behavior. The results are mean (±SD) values from three independent experiments, *p<0.05. Magnification, ×200. F. The results are from four groups that included 10 genes that were present in the up-regulated (ratio, >2.0) genes in the PLC/PRF/5-Sema3A cells and in the down-regulated (ratio, <0.5) genes in HCCLM3-shRNA-Sema3A cells. G. There were also four groups that included 10 genes that were down-regulated by Sema3A overexpression and also up-regulated by Sema3A knockdown. H-I. PLC/PRF/5-Sema3A cells compared with PLC/PRF/5-Mock cells: genes that were up-regulated (76; ratio, 2.0) and down-regulated (94; ratio, 0.5). HCCLM3-shRNA-Sema3A cells compared with HCCLM3-Mock cells: genes that were up-regulated (822; ratio, 2.0) and genes that were down-regulated (671; ratio, 0.5).