微小RNA-10b对人胰腺癌细胞增殖和凋亡的影响  

The effect of microRNA-10b on the proliferation and apoptosis of human pancreatic cancer cells

作　　者：郑岩松[1] 向春晖 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]恩施州中心医院外三科

出　　处：《中华实验外科杂志》2015年第9期2196-2198,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 观察微小RNA（miRNA,miR）-10b对入胰腺癌细胞AsPC-1细胞（AsPC-1）增殖凋亡的影响.方法 重组真核表达载体pPG-CMV-EGFP-miR-10b（实验组）及空载体（对照组）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;细胞计数试剂盒（CCK-8）检测细胞生长曲线;流式细胞仪检测不同组间细胞的凋亡;Western blot检测细胞B细胞淋巴瘤/白血病-2蛋白家族促凋亡调节蛋白（Bim）、髓样细胞白血病-1（MCL-1）蛋白的表达.结果 与对照组（0.002 529±0.000 339）比较,实验组miR-10b表达量（0.020 500±0.007700）增加10倍,差异有统计学意义（P＜0.05）.细胞生长曲线提示,实验组细胞增殖活性较对照组增高24％,差异有统计学意义（P＜0.05）;实验组细胞凋亡率为3.0％,对照组为10.1％,差异有统计学意义（P＜0.05）;实验组细胞Bim蛋白的相对表达水平（0.256 ±0.012）低于对照组（0.652±0.042）,差异有统计学意义（P＜0.05）;实验组细胞MCL-t蛋白的相对表达水平（0.515±0.025）高于对照组（0.154 ±0.068）,差异均有统计学意义（P＜0.05）.结论 miR-10b过表达可调控胰腺癌细胞Bim、MCL-1蛋白的表达,促进细胞增殖,抑制凋亡.Objective To observe the effect of microRNA （miRNA,miR）-10b on the proliferation and apoptosis of human pancreatic cancer cell line AsPC-1.Methods The recombinant eukaryotic expression plasmid vector with miR-10b sequence （pPG-CMV-EGFP-miR-10b） was transfected into AsPC-1 cells with lipofectamine.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 and apoptosis of the cells was evaluated by cell counting kit-8 （CCK-8） assay and flow cytometry.The expression of B-cell lymphoma/Leukemia-2 interacting mediator of cell death （Bim） and myeloid cell leukemia-1 （MCL-1） was detected by Western blotting.Results The expression of miR-10b in the sense group （0.020 500 ±0.007 700） was increased by 10 folds,as compared with the control （0.002 529 ±0.000 339,P ＜0.05）.The ratio of cell proliferation in the experimenal group was 24％ higher than that of control group （P ＜ 0.05）,and the apoptosis ratio in the experimenal group was 3.0％,lower than that of the control （10.1％）.The expression of Bim and MCL-1 protein in the experimental group （0.256 ±0.012 and 0.515 ±0.025） was different with that of control （0.652 ±0.042 and 0.154 ± 0.068,P ＜ 0.05）.Conclusion The overexpression of miR-10b can regulate the expression of Bim and MCL-1 protein,promote proliferation and inhibit the apoptosis of the AsPC-1 cells in vitro.

关 键 词：微小RNA-10b 胰腺癌 增殖 

分 类 号：R737.33[医药卫生—肿瘤]