Figure 2

Ectopic miR-99a expression suppressed migration, invasion and lung colonization. (A) Levels of miR-99a in OEC-M1 and CGHNC9 cells with ectopic miR-99a expression (OEC-M1 miR-99a and CGHNC9 miR-99a) and their corresponding controls with lentiviral non-silencing microRNA (OEC-M1 NS and CGHNC9 NS) were determined using qRT-PCR analysis. MiR-99a expression was normalized against an endogenous control U6. The relative expression of miR-99a was determined by normalizing the expression of miR-99a in cells with ectopic miR-99a expression (miR-99a) to that in control cells (NS). Bar, SE; ***p < 0.001. (B) Cell proliferation was analyzed in OEC-M1 cells with ectopic miR-99a expression (OEC-M1 miR-99a) and controls (OEC-M1 NS). Bar, SE. (C) Cell proliferation was evaluated in CGHNC9 cells with ectopic miR-99a expression (CGHNC9 miR-99a) and controls (CGHNC9 NS). Bar, SE. (D) The relative migration/invasion activity of OEC-M1 cells expressing miR-99a (OEC-M1 miR-99a) and controls (OEC-M1 NS). Bar, SE; * p < 0.05; ***p < 0.001. (E) The relative migration/invasion activity of CGHNC9 cells expressing miR-99a (CGHNC9 miR-99a) and controls (CGHNC9 NS). Bar, SE; * p < 0.05; ***p < 0.001. (F) Representative lung field of nude mice after delivery via tail vein injection of OEC-M1 cells with ectopic miR-99a expression (OEC-M1 miR-99a) or controls (OEC-M1 NS). The white arrowhead indicates tumor nodule in the lung fields. The boxed area in the upper panel is shown at higher magnification in the lower panel. (G) Quantification of colony number/per lung section in mice injected with miR-99a expressing OEC-M1 (OEC-M1 miR-99a) or controls (OEC-M1 NS). Bar, SE; *p < 0.05.