Figure 7. Stromal cells, through production of Shh, support cancer cells growth in non adherent conditions.

(A) CAL62 cells (50 cells/well) were grown on ultra-low-attachment 96-well in low serum medium alone or together with MSC or IMR90 cells (20 cells/wells). Spheroids were photographed after 7 and 14 days. Cancer cells, supported by stromal cells, formed larger spheroids and this ability was impaired by cyclopamine (10 mM) treatment as well as by 5E1 blocking antibody (2.5 μg/ml). Dependence of stromal cells effect on Shh production was tested by performing Gli luciferase assay (B), Q-RT-PCR (C) and Matrigel assay (D) on un-stimulated and MSC stimulated CAL62 cells with and without 5E1 blocking antibody. Error bars represent standard deviations of experimental triplicates. ^**p = ≤ 0.01; ^*p = ≤ 0.0.