3.

HS promotes tumor-initiating cell traits and invasion capacity. (A) Phase-contrast pictures of tumor spheres formed by HS treated CT-26 cells, grown in serum-free sphere medium on ultra-low attachment surface plates (scale bar, 200 μm). (B, C) Quantification of tumor spheres formed by CT-26 cells. The mean number of spheres per 5,000 cells (B) and the mean diameter of spheres (C) are calculated in triplicate samples. (D) Matrigel pre-coated Transwell assays were performed to examine the invasion ability of CT-26 and SW620 cell 3 days after HS treatment, CT-26 (24hrs), SW-620 (36hrs). Werstern blot assay was performed with cell lysate and Tissue culture Condition Media (TCM). Gelatin zymography assay was performed with TCM. The quantification value was labeled above the bands. P values were determined by ANOVA test. * P < 0.05, ** P < 0.01. All data were from three independent experiments.