微小RNA-25靶向大型肿瘤抑制因子2对结肠癌细胞增殖及凋亡的影响  

MicroRNA-25 regulates proliferation and apoptosis of colorectal cancer cells by targeting large tumor suppressor homolog 2

作　　者：吴万庆[1] 崔小兵[1] 傅聿铭[1] 吴刚[2] 陈广法 

机构地区：[1]郑州大学第五附属医院普外科,450052 [2]河南省人民医院普外科,郑州450003 [3]河南省开封市儿童医院儿科,475000

出　　处：《中华实验外科杂志》2017年第7期1152-1155,共4页Chinese Journal of Experimental Surgery

基　　金：河南省自然科学基金（面上项目）（162300410292）

摘　　要：目的观察微小RNA（miRNA，miR）-25靶向大型肿瘤抑制因子2（LATS2）对结肠癌细胞增殖及凋亡的影响。方法选取人结肠癌细胞HT29，转染Neg-miR、pre-miR-25、anti-miR-25，采用反转录-聚合酶链反应（RT-PCR）法检测miR-25表达，Western blot检测LATS2蛋白表达，噻唑蓝（MTT）法检测细胞增殖活性，流式细胞仪检测细胞凋亡。双荧光素酶报告基因实验验证miR-25与LATS2的靶向关系。结果与Neg-miR组比较，pre-miR-25组miR-25表达上调（0.37±0.08比0.78±0.14）（t=3.871，P=0.028），LATS2蛋白表达降低（157.32±32.11比56.47±11.35） （t=4.012，P=0.026）；anti-miR-25组miR-25表达降低（0.37±0.08比0.11±0.04）（t=4.275，P=0.025），LATS2蛋白表达增高（157.32±32.11比382.74±55.28）（t=4.663，P=0.021）。与Neg-miR组比较，pre-miR-25组细胞增殖活性增高（0.37±0.04比0.48±0.02，0.62±0.05比0.90±0.02，0.89±0.03比1.10±0.04）（t=2.998、3.372、3.012，P=0.045、0.035、0.041），同时凋亡率降低[（8.52±1.11）%比（2.94±0.81）%，t=4.425，P=0.018]；anti-miR-25组细胞增殖活性降低（0.37±0.04比0.30±0.03，0.62±0.05比0.42±0.04，0.89±0.03比0.50±0.05）（t=2.895、3.237、4.002，P=0.048、0.038、0.021），同时凋亡率降低[（8.52±1.11）%比（15.01±2.53）%，t=4.518，P=0.016]。miRNA靶基因预测软件检测结果显示，miR-25能作用于LATS2 3’端非编码区域（3’UTR）。与转染Neg-miR比较，共转染pre-miR-25与LATS2-wt可使HT29荧光素酶活性降低（t=3.285，P=0.033）。结论miR-25可通过靶向LATS2调控结肠癌细胞增殖及凋亡。Objective To evaluate the regulatory effect of microRNA （miRNA, miR）-25 on proliferation and apoptosis of colorectal cancer cells by targeting large tumor suppressor homolog 2 （LATS2）.Methods The colorectal cancer cell line HT29 was collected, and respectively transfected with Neg-miR, pre-miR-25 and anti-miR-25 separately. The miR-25 expression was detected by reverse transcriptase-polymerase chain reaction （RT-PCR）. The LATS2 protein expression was detected by Western blotting. The cell proliferation activity was detected by methyl thiazol tetrazolium （MTT） method. The cell apoptosis was examined by flow cytometry. The targeted relationship between miR-25 and LATS2 was tested by dual-luciferase report gene experiment.Results As compared with Neg-miR group, the miR-25 expression in pre-miR-25 group was significantly increased （0.37±0.08 vs. 0.78±0.14） （t=3.871, P=0.028）, and the LATS2 protein expression significantly decreased （157.32±32.11 vs. 56.47±11.35） （t=4.012, P=0.026）. The miR-25 expression in anti-miR-25 group was significantly decreased （0.37±0.08 vs. 0.11±0.04） （t=4.275, P=0.025）, and the LATS2 protein expression significantly increased （157.32±32.11 vs. 382.74±55.28） （t=4.663, P=0.021）. As compared with Neg-miR group, the proliferation activity in pre-miR-25 group was significantly increased （0.37±0.04 vs. 0.48±0.02, 0.62±0.05 vs. 0.90±0.02, and 0.89±0.03 vs. 1.10±0.04） （t=2.998, 3.372, 3.012, P=0.045, 0.035, 0.041）, and the apoptosis rate was significantly decreased [（8.52±1.11）% vs. （2.94±0.81）%, t=4.425, P=0.018]. The proliferation activity in anti-miR-25 group was significantly decreased （0.37±0.04 vs. 0.30±0.03, 0.62±0.05 vs. 0.42±0.04, and 0.89±0.03 vs. 0.50±0.05） （t=2.895, 3.237, 4.002, P=0.048, 0.038, 0.021）, and the apoptosis rate significantly increased [（8.52±1.11）% vs. （15.01±2.53）%, t=4.518, P=0.016]. The miRNA target genes prediction software test results showed that miR-25 could act

关 键 词：微小RNA-25 大型肿瘤抑制因子2 结肠癌 增殖 

分 类 号：R735.35[医药卫生—肿瘤]