Objective: To investigate the effect of microRNA-10b (miR-10b) expression alteration on the biological behavior of human pancreatic cancer ASPC-1 cells. Methods: ASPC-1 cells were transfected with the eukaryotic expression plasmid vector containing pre-miR-10b and antisense miR-10b or empty vector by lipofectamine, respectively, and the transfection efficiency was measured with fluorescence microscopy. Using the non-transfected ASPC-1 cells as blank control, the miR-10b and RhoC protein expression of the transfected cells in each group were detected, and their growth, apoptosis and invasve ability were also examined. Results: The transfection efficiencies of each group were about 50%-70% 48 h after transfection. By comparison with blank control group, cells in pre-miR-10b transfection group showed increased expression of miR-10b and RhoC protein, decreased apoptosis, and enhanced proliferative and invasion ability (all P<0.05), while the results of above indexes in antisense miR-10b transfection group showed the opposite changes to those in pre-miR-10b transfection group (all P<0.05). Bat there were no significant differences in all determined parameters between empty vector transfection group and blank control group (all P>0.05). Conclusion: Up-regulation of miR-10b in ASPC-1 cells can promote their growth and invasive ability with reduced apoptosis, and these effects may probably be associated with its action on regulation of RhoC expression.