Figure 2

TAMs promote proliferation, mobility and invasiveness of PC-3 cells and promote angiogenesis.

(a) Proliferation of PC-3 cells. PC-3 cells were exposed to CM from M0, M1, M2 and TAMs or 1640 medium as control for 72 h. Viability of PC-3 cells was measured by MTT assay. (b) Mobility of PC-3 cells. A line of PC-3 cells was scraped away in each well using a pipette tip after 6 h of serum starvation. Subsequently, cells were treated with CM from M0, M1, M2 and TAMs for 24 h. Migrated cells were observed from three randomly chosen fields (original magnification, 100×) and the number of migrated cells was quantified by manual counting. (c) Invasiveness of PC-3 cells. PC-3 cells were loaded into the upper compartments and then placed into 24-well culture dishes containing different CM from M0, M1, M2 and TAMs, or RPMI 1640 medium as control. After 24 h of incubation at 37 °C, cells that migrated to the bottom of the membrane were stained with hematoxylin-eosin and counted using an inverted microscope (original magnification, 100×). (d) Tube-like structure formation in HUVEC. HUVEC were seeded to the matrigel-coated plates, followed by addition of CM from M0, M1, M2 or TAMs, respectively. The effects on the morphogenesis of endothelial cells were recorded after 5 h with an inverted microscope equipped with CCD optics and a digital analysis system. Results were quantified by measuring the joint or vessel numbers in the field (original magnification, 100×). *P < 0.05; **P < 0.01; ***P < 0.001.