Fig. 3.

Effect of PRSS3 on the growth properties of human HCC cells. HCC HepG2 and PLC/PRF/5 cell lines with overexpression of PRSS3, and SNU-387 cell line with knockdown of PRSS3 expression were established to evaluate the potential roles of PRSS3 in HCC development. a, b MTT assays showed the viability of HCC HepG2 and PLC/PRF/5 cells with PRSS3 overexpression (PRSS3) or vector control (Vector) (a), or SNU-387 cells transfected with either siRNA against PRSS3 (siPRSS3–2) [20] or RNAi Negative Control Duplex (siNC) (b). c, d. Colony formation in soft agar for 2 weeks by HCC HepG2 and PLC/PRF/5 cells with PRSS3 overexpression, or control (C), or SNU-387 cells transfected with siPRSS3–2 or siNC (d). Left panel: representative image; Right panel: quantitative analysis. e, f FACS analysis of cell cycle distribution of HepG2 and PLC/PRF/5 cells with or without ectopic expression of PRSS3 (E), or SNU-387 cells transfected with siPRSS3–2 or siNC (f). Left panel: representative FACS histograms with the percentage of cells in each cell cycle phase (Left red peak: G0/G1; right red peak: G2/M; hatched peak: S; coefficient of variation of G1 peak: % CV); right panel: quantitative graphs. g Western blotting analysis of the levels of PRSS3, cyclin D1, CDK4, cyclin E1, and CDK2 in the transfected HCC cell lines. The experiments were repeated at least three times, and the results were presented as the mean ± SD, *p < 0.05, versus control