国际肿瘤学杂志

国际肿瘤学杂志 ›› 2016, Vol. 43 ›› Issue (11): 801-805.doi: 10.3760/cma.j.issn.1673-422X.2016.11.001

• 论著 •    下一篇

微小RNA-143抑制食管癌细胞株ECA109的增殖、迁移和侵袭

吴文学，刘万平，刘合波，陈洁，孙艳燕，邹传涛，李艳梅   

  1. 441300 湖北医药学院附属随州医院肿瘤科
  • 收稿日期:2016-03-20 出版日期:2016-11-08 发布日期:2016-11-02
  • 通讯作者: 李艳梅，Email: 410756836@qq.com E-mail:410756836@qq.com

MicroRNA-143 inhibits proliferation and migration as well as invasion in esophageal cancer cell line ECA109

Wu Wenxue, Liu Wanping, Liu Hebo, Chen Jie, Sun Yanyan, Zou Chuantao, Li Yanmei   

  1. Department of Oncology, Suizhou Hospital, Hubei University of Medicine, Suizhou 441300, China
  • Received:2016-03-20 Online:2016-11-08 Published:2016-11-02
  • Contact: Li Yanmei E-mail:410756836@qq.com

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摘要： 目的 研究微小RNA143(miR143)在食管癌细胞株ECA109中的作用。方法ECA109细胞分为转染阴性对照组、miR143模拟剂组和miR143抑制剂组，四甲基偶氮唑蓝（MTT）法检测各组细胞的增殖；Annexin VFITC/PI凋亡检测试剂盒检测细胞凋亡；Transwell实验检测各组细胞的迁移和侵袭能力；实时荧光定量PCR和Western blotting分别检测Kras mRNA及蛋白表达水平。结果转染3 d和4 d后，与阴性对照组相比(吸光度值分别为0.90±0.02和1.09±0.07)，miR143模拟剂组ECA109细胞增殖受抑制(吸光度值分别为0.66±0.05和0.80±0.04)，而miR143抑制剂组细胞增殖增加（吸光度值分别为1.13±0.09和1.51±0.08），各组之间差异有统计学意义（F=49.16，P=0.000；F=100.34，P=0.000）。阴性对照组、miR143模拟剂和抑制剂组的早期凋亡率分别为3.42%±0.72%、11.63%±1.15%和0.94%±0.10%，各组之间差异有统计学意义（F=151.61，P=0.000）。与阴性对照组相比（迁移细胞数336±13，侵袭细胞数147±16），miR143模拟剂组的细胞迁移（148±16）和侵袭（75±10）能力降低，而miR143抑制剂组的细胞迁移（510±14）和侵袭（238±16）能力增加，各组之间差异有统计学意义（F=470.99，P=0.000；F=90.04，P=0.000）。相对于阴性对照组(1.00±0.00)，miR143模拟剂抑制ECA109细胞中Kras mRNA（0.22±0.08）及蛋白（0.46±0.08）表达，而miR143抑制剂上调Kras mRNA（1.55±0.12）及蛋白（1.33±0.05）表达，各组之间差异有统计意义（F=131.36，P=0.000；F=88.17，P=0.000）。结论miR143在食管癌细胞株ECA109中发挥抑癌基因作用，下调Kras蛋白表达可能是其发挥抑癌作用的机制之一。

关键词: 微RNAs, 食管肿瘤, 基因, 肿瘤抑制

Abstract: Objective To investigate the functions of microRNA143 (miR143) in esophageal cancer cell line ECA109. MethodsECA109 cells were transfected with negative control (NC), miR143 mimics or miR143 inhibitors. 3(4,5dimethyl2thiazolyl)2,5diphenyl2Htetrazolium bromide (MTT) assay was performed to evaluate the growth of ECA109 cells after transfection. Annexin VFITC/PI apoptosis test kit was used to detect early apoptosis rate in ECA109 cells. Transwell migration and invasion assays were conducted to compare the migration and invasion capacity of ECA109 among different groups. Realtime PCR and Western blotting were used to analyze the mRNA and protein alteration after transfection. ResultsThree and four days after transfection, compared with NC (absorbance value: 0.90±0.02 and 1.09±0.07), miR143 mimics inhibited ECA109 cell proliferation (absorbance value: 0.66±0.05 and 0.80±0.04), while miR143 inhibitors promoted cell proliferation (absorbance value: 1.13±0.09 and 1.51±0.08), with statistical significances (F=49.16, P=0.000; F=100.34, P=0.000). Earlystage apoptosis rates of ECA109 transfected with NC, miR143 mimics and miR143 inhibitors were 3.42%±0.72%, 11.63%±1.15% and 0.94%±0.10%, respectively, with statistical significance (F=151.61, P=0.000). Meanwhile, compared with NC (migration cell number: 336±13, invasion cell number: 147±16), miR143 mimics inhibited cell migration (148±16) and invasion (75±10), while miR143 inhibitors promoted cell migration (510±14) and invasion (238±16), with statistical significances (F=470.99, P=0.000; F=90.04, P=0.000). Compared with NC (1.00±0.00), miR143 mimics downregulated mRNA (relative expression level 0.22±0.08) and protein expression (relative expression level 0.46±0.08) of Kras, whereas miR143 inhibitors upregulated mRNA (1.55±0.12) and protein expression (1.33±0.05) of Kras (F=131.36, P=0.000; F=88.17, P=0.000). ConclusionmiR143 functions as a tumor suppressor in esophageal cancer cell line ECA109, probably by downregulating Kras expression.

Key words: MicroRNAs, Esophageal neoplasms, Genes, tumor suppressor