Figure 2.

Serglycin suppression reduces CSC-like properties of S18 cells in vitro and in vivo. S18 cells were stably transfected with one of two serglycin shRNAs (KD1, KD2) or scrambled shRNA. (a) The expression of serglycin was detected by quantitative real-time PCR and immunoblotting in S18 and S26 stable cell lines. Serglycin protein levels were not changed in whole-cell lysates (WCL), but secreted serglycin protein was significantly reduced in CM after serglycin knockdown. (b) The change in cellular morphology of stably transfected cells (40 ×) . (c) Serglycin knockdown markedly decreased the expression of stem cell-associated genes. Data represent the average±S.D., n=3; *P<0.05 (d) Serglycin suppression reduced the size and number of spheres generated by S18 stable cells. Data represent the average±S.D., n=3; *P<0.05 relative to the scrambled controls. (e) S18 scrambled cells or S18 SG KD2 cells were subcutaneously inoculated into nude mice and were observed twice per week for 30 days. IHC staining of serglycin in S18 scrambled tumor tissues and in S18 SG KD2 tumor tissues (bottom panel, 200 ×)