microRNA-10b表达变化对人胰腺癌细胞生物学行为的影响

目的:探讨microRNA-10b(miR-10b)的表达变化对人胰腺癌细胞ASPC-1生物学行为的影响。方法:胰腺癌细胞ASPC-1细胞株分别以脂质体法转染正义、反义miR-10b真核表达载体和空载体,用荧光显微镜观察转染效率。以未转染的ASPC-1细胞为空白对照,检测各转染组miR-10b和RhoC蛋白表达,并检测各组细胞生长、凋亡及侵袭能力。结果:各组转染48 h后,转染效率达50%~70%;与空白对照组比较,正义miR-10b转染组细胞miR-10b表达和RhoC蛋白表达量明显增加,凋亡率明显降低,生长活性和侵袭能力明显增强(均P<0.05);而反义miR-10b转染组细胞以上检测结果与正义miR-10b转染组细胞呈相反改变(均P<0.05);空载体转染组所有指标与空白对照组间无明显差异(均P>0.05)。结论:上调miR-10b的表达,能促进胰腺癌细胞ASPC-1的生长、侵袭并抑制凋亡,该作用可能与其调控RhoC表达有关。

Effect of microRNA-10b expression alteration on biological behavior of human pancreatic cancer cells

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.