微小RNA-16调控卵巢上皮性癌细胞增殖、侵袭及凋亡的体外研究  

MicroRNA-16 regulates the proliferation,invasion and apoptosis of ovarian epithelial carcinoma cells in vitro

作　　者：唐蕊[1] 崔竹梅[1] 娄艳辉[1] 

机构地区：[1]青岛大学医学院附属医院妇科,266001

出　　处：《中华妇产科杂志》2012年第11期846-850,共5页Chinese Journal of Obstetrics and Gynecology

摘　　要：目的 探讨微小RNA-16（miR-16）调控卵巢上皮性癌（卵巢癌）细胞增殖、侵袭及凋亡的作用及机制.方法 脂质体介导miR-16成熟体模拟物及相应的阴性对照片段转染人卵巢癌细胞SKOV-3,分别作为转染组、阴性对照组.实时荧光定量PCR技术检测各组细胞中miR-16的表达,蛋白印迹法检测血管内皮生长因子（VEGF）、基质金属蛋白酶2（MMP-2）、凋亡抑制蛋白bcl-2的表达,采用四甲基偶氮唑蓝法、5-乙炔基-2’脱氧尿苷（EdU）标记法测定细胞的增殖能力,transwell穿膜小室侵袭实验检测细胞的侵袭能力,流式细胞仪检测血清饥饿及低氧条件下细胞的凋亡情况.结果（1）转染组、阴性对照组SKOV-3细胞中miR-16的表达量分别为125.93±15.30、 0.78±0.16,转染组明显高于阴性对照组,两组比较,差异有统计学意义（P＜0.01）.（2）转染组、阴性对照组SKOV-3细胞中VEGF蛋白的相对表达量为0.58±0.05、1.22±0.03,MMP-2蛋白的相对表达量为0.63±0.03、1.16±0.03,bcl-2蛋白的相对表达量为0.52±0.03、1.19±0.05,转染组各蛋白的相对表达量均明显低于阴性对照组,分别比较,差异均有统计学意义（P＜0.01）.（3） 转染后第4天,转染组、阴性对照组细胞的增殖抑制率分别为（37.2±6.2）％、（3.6±3.2）％,两组比较,差异有统计学意义 （P=0.001）.（4）转染组、阴性对照组细胞的增殖率分别为（12.3±0.8）％、（23.4±1.8）％,转染组明显低于阴性对照组,两组比较,差异有统计学意义（P＜0.05）.（5）转染组穿过聚碳酸酯膜的细胞数为（6±3）个,明显少于阴性对照组的（40±9）个,两组比较,差异有统计学意义（P＜0.01）.（6）血清饥饿及低氧条件培养24 h后,转染组细胞的早期凋亡率[（16.9±2.1）％]、晚期凋亡率[（13.4±3.3）％]均明显高于阴性对照组[分别为 （10.3±1.7）％、（3.2±1.8）％],分别比较,差异均有统计学意义（P＜0.01）;培Objective To study the role and mechanism of microRNA-16（miR-16）in the proliferation,invasion and apoptosis of ovarian epithelial carcinoma cells in vitro.Methods The SKOV-3 cells were transfected with miR-16 mimics or negative control RNA（NC）by lipofectamine 2000.The expression of miR-16 was detected by real-time reverse transcription（RT）-PCR in SKOV-3 cells,and western blot was used to detect the expression of vascular endothelial growth factor（VEGF）,matrix metalloproteinase-2（MMP-2）and bcl-2 protein.Methyl thiazolyl tetrazolium（MTT）,5-ethynyl-2＇-deoxyuridine（EdU）and transwell assay were used to determine the proliferation and invasion abilities.And the rate of apoptotic cell was detected by flow cytometry method.Results（l）The expression level of miR16 in the transfection cells group was significantly higher than that in NC group（125.93 ± 15.30 versus 0.78 ± 0.16,P 〈 0.01）.（2）The rclative expression level of VEGF protein in transfection cells,NC and blank control group was 0.58 ± 0.05,1.22 ± 0.03,1.20 ± 0.03,MMP-2 protein was 0.63 ± 0.03,1.16 ±0.03,1.21 ± 0.03,and bel-2 protein 0.52 ± 0.03,1.19 ± 0.05,1.28 ± 0.06,respectively.The level of VEGF,MMP-2 and bcl-2 protein in the transfection group were lower than those in other control groups,and there were significantly differences among them（all P 〈0.01）.（3）After transfected 4 days,the inhibition rate of cell proliferation in the transfection group was dramatically higher than that in NC group[（37.2 ±6.2）% versus（3.6 ± 3.2）%,P =0.001].（4）The percentage rate of proliferative cells in the transfection,NC and blank control group was（12.3 ± 0.8）%,（23.4 ± 1.8）%,（31.1 ± 4.9）%.And it was lower in the transfection group（P 〈 0.05）.（5）Decreased cells via the transwell member in the transfection group（6 ± 3）were detected as compared with NC group（40 ± 9）and blank control group （48 ± 8,P 〈 0.01）.（6）Twenty-four hours after cultured in serum starvation an

关 键 词：微RNAs 卵巢肿瘤 细胞增殖 肿瘤浸润 细胞凋亡 原癌基因蛋白质 c-bcl-2 体外研究 

分 类 号：R737.31[医药卫生—肿瘤]