Effect of transfection of microRNA-10b and antisense microRNA-10b on the expression of RhoC in human pancreatic cancer cell line AsPC-1

Abstract

Objective To observe the effect of microRNA (miR)-10b on the expression of RhoC in human pancreatic cancer cell line AsPC-1.Methods The recombinant eukaryotic expression plasmid vector with miR-10b sequence [pPG-CMV-enhanced green fluorescent protein (EGFP)-miR-10b] or the antisense sequence was transfected into human pancreatic cancer cell line (AsPC-1) with lipofectamine,and the efficiency of the instant expression was confirmed by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) and fluorescence microscope.The difference of the migration and invasion of the cells was evaluated by matrigel assay,and the expression of RhoC was evaluated by Western blotting.Results The expression of miR-10b in the sense group was increased by 10 folds,and in antisense group,the expression was decreased to 14％ compare to the control (P ＜ 0.05).The expression of RhoC protein in the sense group was (95.7 ± 8.5) unit,and that in the antisense group (28.5 ± 5.7) unit (P ＜0.05).As compared to the antisense group,the number of migrating and penerating cells was decreased from (85 ±7) and (35 ±9) to (32±5) and (8 ±3) in the sense group (P＜0.05) Conclusion miR10b can up-regulate the expression of RhoC,and promote the migration and invasion of the AsPC-1 cells. Key words: MicroRNA-10b; Pancreatic cancer ; Migration ; Invasion