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目的 观察微小RNA(miR)-10b沉默对人胰腺癌细胞AsPC-1细胞(以下简称AsPC-1)表达过氧化物酶增殖体激活受体-α蛋白(PPAR-α)的影响.方法 以脂质体转染法将重组反义真核表达载体pPG-CMV-EGFP-miR-10b转染AsPC-1细胞株,实时定量聚合酶链反应(Real-time PCR)和荧光显微镜检测载体瞬时表达的效率,细胞增殖实验评价不同组间细胞增殖能力的差异;Western blot 检测细胞PPAR-α蛋白的表达.结果 转染反义组miR-10b减少至对照组的14％.细胞增殖减少至对照组的36％,差异有统计学意义(P＜0.05).细胞PPAR-α蛋白的表达水平:反义组1 230.5±110.4,对照组2 398.1±156.3和2 603.7±227.5,反义组与对照组差异均有统计学意义(P＜0.05);细胞凋亡实验:反义组凋亡率为19.4％,对照组凋亡率为10.1％和11.5％.结果提示反义组细胞的凋亡率增高,差异有统计学意义(P＜0.05).结论 miR-10b沉默可抑制胰腺癌细胞PPAR-α蛋白的表达,并抑制胰腺癌细胞增殖,促进细胞凋亡.

Objective To observe the effect of silencing microRNA-10b on the expression of peroxisome proliferator activated receptor-α(PPAR-α) in human pancreatic cancer cell line AsPC-1.Methods The recombinant eukaryotic expression plasmid vector with miR-10b antisense sequence was transfected into AsPC-1 cells with lipofectamine,and the efficiency of the instant expression was confirmed by real-time quantitative polymerase chain reaction (Real-time PCR) and fluorescence microscope.The difference of the proliferation of the cells was measured by proliferation assay,and the expression of PPAR-α was detected by Western blotting.Results The expression of miR-10b in the antisense groups was decreased to 14％ as compared with the control group (P ＜ 0.05).The cell proliferation in the antisense groups was decreased to 36％ as compared with the control group (P ＜ 0.05).The expression of PPAR-α protein in the antisense groups was 1 230.5 ± 110.4,and that in the control groups was 2 398.1 ± 156.3 and 2 603.7 ± 227.5 (P ＜0.05).The apoptosis ratio in the antisense group was 19.4％,and that in the control groups was 10.1％ and 11.5 ％ (P ＜ 0.05).Conclusion The silencing of miR-10b can inhibit the expression of PPAR-α and the cell proliferation,and promote the apoptosis of the ASPC-1 cells.