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. Author manuscript; available in PMC: 2014 Jan 13.
Published in final edited form as: Genes Chromosomes Cancer. 2012 Oct 29;52(3):225–236. doi: 10.1002/gcc.22022

Figure 2.

Figure 2

Knockdown of SULF2 suppresses the antitumor effect of OKN-007 in Huh7 cells. (A) Both SULF2 mRNA and protein were significantly downregulated by stable transfection with SULF2 shRNAs as assessed by qRT-PCR and caspase. (B) Knockdown of SULF2 resulted in an increased percent apoptosis in untreated Huh7 SULF2 shRNA cells and a lower fold increase in apoptosis after OKN-007 treatment (3.4-fold). In contrast, untreated control Huh7 Scr shRNA cells exhibited a lower percent apoptosis and were more sensitive to OKN-007-induced apoptosis, with a 5.2-fold increase in apoptosis after OKN-007 treatment. (C) Cells transfected with control scrambled shRNA exhibited a significant increase after treatment with OKN-007, whereas cells transfected with SULF2 shRNAs exhibited no significant difference in caspase 3/7 activity after OKN-007 treatment. The Caspase-Glo® 3/7 Assay kit was used and the unit of the results is RLU. (D) Knockdown of SULF2 decreased the basal rate of cell proliferation, and also substantially decreased the effect of OKN-007 on proliferation of Huh7 cells. (E) Basal cell viability of Huh7 cells was not significantly altered by transfection with shRNAs targeting SULF2. OKN-007 treatment of the SULF2 expressing control cells led to a significant decrease in cell viability, whereas the shRNA-transfected cells with suppressed SULF2 exhibited no change in cell viability on treatment with OKN-007. (F) OKN-007 treatment for 48 h substantially inhibited migration of control Huh7 Scr shRNA cells by 47%; in contrast, Huh7 SULF2 shRNA cells exhibited a much lower 17% inhibition of migration in response to OKN-007. (G) The inhibitory effect of OKN-007 on migration ability of SNU387 cells was decreased by silencing SULF2 expression with shRNAs targeting SULF2.