微小RNA-1Ob沉默抑制胰腺癌细胞表达过氧化物酶增殖体激活受体-α  

Effect of microRNA-10b silencing on expression of peroxisome proliferator activated receptor-a in human pancreatic cancer cell line

作　　者：郑岩松[1] 林俊[2] 林振吕[2] 石铮[1] 

机构地区：[1]福建医科大学附属第一医院肝胆胰外科,福州350005 [2]福建医科大学附属第一医院急诊外科,福州350005

出　　处：《中华实验外科杂志》2014年第10期2175-2177,共3页Chinese Journal of Experimental Surgery

摘　　要：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.

关 键 词：胰腺癌微小RNA-10b 过氧化物酶增殖体激活受体-α 

分 类 号：R735.9[医药卫生—肿瘤]