上调miR-200b对人喉癌Hep-2细胞增殖、迁移和侵袭能力的影响  

Up-regulation of miR-200b may inhibit epithelial mesenchymal transition process to prevent the proliferation,migration,and invasion of human laryngeal cancer Hep-2 cells

作　　者：李国彬[1] 张占成[1] 王新颜[1] LI Guobin;ZHANG Zhancheng;WANG Xinyan(Department of Otolaryngology, Fourth People＇s Hospital of Jinan, Jinan 250031, Shandong, China)

机构地区：[1]济南市第四人民医院耳鼻喉科,山东济南250031

出　　处：《山东大学耳鼻喉眼学报》2018年第4期53-57,共5页Journal of Otolaryngology and Ophthalmology of Shandong University

摘　　要：目的探讨上调miR-200b对人喉癌Hep-2细胞增殖、迁移和侵袭能力的影响。方法培养人喉癌Hep-2细胞,随机分为miR-200b模拟物组、miR-对照序列组和空白对照组,利用实时荧光定量PCR技术检测各组细胞中miR-200b表达,MTT法检测细胞增殖能力,Transwell法检测细胞迁移和侵袭能力,Western blotting法检测各组细胞中E-cadherin、N-cadherin、β-catenin蛋白表达。结果与空白对照组和miR-对照序列组相比,miR-200b模拟物组细胞中miR-200b相对表达量显著升高,差异有统计学意义(F=70.766,P<0.001);与空白对照组[(0.22±0.05)、(0.45±0.07)、(0.59±0.06)、(0.72±0.08)和(0.85±0.07)]和miR-对照序列组[(0.24±0.06)、(0.46±0.08)、(0.61±0.07)、(0.70±0.05)和(0.84±0.06)]相比,miR-200b模拟物组细胞24、48、72、96 h时吸光度A值[(0.21±0.04)、(0.29±0.06)、(0.40±0.04)、(0.53±0.07)和(0.58±0.05)]均降低,差异均有统计学意义(F=0.134,P=0.876;F=6.449,P=0.010;F=37.299,P<0.001;F=5.352,P=0.018;F=29.921,P<0.001);与空白对照组[(140.2±2.3)、(127.9±6.0)]和miR-对照序列组[(141.0±1.2)、(130.4±7.4)]相比,miR-200b模拟物组迁移细胞数和侵袭细胞数[(98.5±2.4)、(90.9±2.8)]均减少,差异均有统计学意义(F=845.523,P<0.001;F=88.859,P<0.001);与空白对照组[(0.35±0.07)、(0.54±0.10)、(0.76±0.12)]和miR-对照序列组[(0.24±0.03)、(0.60±0.14)、(0.65±0.24)]相比,miR-200b模拟物组细胞中E-cadherin蛋白相对表达量(0.54±0.14)升高,而N-cadherin、β-catenin蛋白相对表达量[(0.31±0.10)、(0.35±0.06)]降低,差异均有统计学意义(F=17.287,P<0.001;F=10.083,P=0.002;F=10.434,P=0.001)。结论上调喉癌细胞中miR-200b基因表达可减少细胞增殖,抑制细胞迁移和侵袭能力,其机制可能与抑制上皮-间质转化过程有关。Objective We investigated the effects of up-regulation of miR-200 b on the proliferation,migration,and invasion of human laryngeal cancer Hep-2 cells.Methods Human laryngeal carcinoma Hep-2 cells were cultured and randomly divided into the miR-200 b mimic group,miR-control sequence group,and blank control group.The expression of miR-200 b was detected by real-time fluorescence quantitative PCR.The MTT assay was used to detect cell proliferation.The Transwell method was used to detect cell migration and invasion.The expression of E-cadherin,N-cadherin,and β-catenin proteins was detected by western blotting.Results Compared to the blank control group and miR-control sequence group,the relative expression level of miR-200 b in the miR-200 b mimic group was significantly increased（F=70.766,P〈0.001）.Compared to the blank control group [（0.22±0.05）,（0.45±0.07）,（0.59±0.06）,（0.72±0.08）,and（0.85±0.07） ]and miR-control sequence group [（0.24±0.06）,（0.46±0.08）,（0.61±0.07）,（0.70±0.05）,and（0.84±0.06） ],the absorbance A values at 24,48,72,and 96 h in the miR-200 b mimic group [（0.21±0.04）,（0.29±0.06）,（0.40±0.04）,（0.53±0.07）,and（0.58±0.05）] were decreased and the differences were statistically significant（F=0.134,6.449,37.299,5.352,and 29.921,P=0.876,0.010,0.000,0.018,and 0.000,P〈0.05）.Compared to the blank control group [（140.2±2.3）,（127.9±6.0） ]and miR-control sequence group [（141.0±1.2）,（130.4±7.4） ],the number of migrating cells and number of invasive cells in the miR-200 b mimic group [（98.5±2.4）,（90.9±2.8） ]were decreased and the differences were statistically significant（F=845.523,88.859,P both 0.001）.Compared to the blank control group [（0.35±0.07）,（0.54±0.10）,（0.76±0.12） ] and miR-control sequence group [（0.24±0.03）,（0.60±0.14）,（0.65±0.24） ],the relative expression level of E-cadherin protein in miR-200 b mimic group cells（0.54±0.14） was increased,whereas the relati

关 键 词：喉癌 miR-200b 细胞增殖 细胞侵袭 上皮-间质转化 

分 类 号：R739.6[医药卫生—肿瘤]