摘要
目的观察微小RNA（miRNA，miR）-21对人肿瘤细胞生长和侵袭能力的影响。方法人喉癌Hep-2细胞分为3组：空白对照组（Con-A）、空载对照组（Con-B）和反义寡核苷酸（AS-miR-21），其中，AS-miR-21组以miR-21反义寡核苷酸组转染处理。测定荧光素酶活性验证3组细胞中miR-21的表达，分别采用噻唑蓝（MTT）法和克隆形成实验检测癌细胞生长和克隆形成能力，以Transwell方法检测细胞的侵袭能力。结果荧光素酶活性结果显示，分别与Con—A组和Con-B组比较，转染AS-miR-21组Hep-2细胞miR-21表达水平降低；MTT结果显示，72h Con-A组、Con-B组和AS-miR-21组吸光度（A）值分别为0．800±0．035、0．791±0．024和0．447±0．014（P〈0．05）；平板克隆结果显示，Con-A组、Con-B组和AS-miR-21组克隆形成数分别为104±8、96±4和44±3（P〈0．05）；Transwell结果显示，Con-A、Con-B和AS-miR-21组穿膜细胞数分别为154±8、141±4和70±9（P〈0．05）。结论miR-21在人喉癌细胞增殖、侵袭中发挥重要作用。
Objective To study the effects of microRNA （miRNA, miR） -21 on growth and in- vasion of human laryngopharynx cancer Hep - 2 cells. Methods The Hep - 2 cells were classified into three groups： blank control group （ Con - A）, the empty plasmid group （ Con - B ） and antisense mill - 21 group （As -miR- 21 ）. The transfection reagent oligofectamine was mixed with antisense miRNA -21 （ AS - miR - 21 ） and nonsense oligodeoxyribonucleotides （ ODN）, respectively, and then, they were added into the medium of Hep -2 cells as AS -mill -21 treatment group and nonsense ODN treatment group, respectively. The miR -21 knocking down effects were examined by luciferase activity assay. The growth of cancer cells was determined by methyl thiazol tetrazolium （MTT） assay and clone formation assay, and the invasion ability was studied by Transwell assay. Results Luciferase intensity in As - mill - 21 treated cancer cells was significantly suppressed as compared with that in the control group and nonsense ODN treatment group （P 〈 0. 05 ）. After cancer cells were transfected by As - miR -21, the results of MTr assay showed that the absorbance （A） values in Con - A, Con - B and As - miR -21 groups at 72 h were 0. 800 ±0. 035, 0. 791 ±0. 024 and 0. 447 ±0. 014 respectively （P 〈0. 05） ; the results of colony formation assay showed that clones in Con - A, Con - B and As - miR - 21 groups were 104 ± 8, 96 ± 4 and 44 ± 3 （P 〈 0. 05）. The results of Transwell showed that the number of cells penetrating membrane in Con - A, Con - B and FoxM1 groups was 154 ± 8, 141 ± 4 and 70 ± 9, respectively （ P 〈 0. 05 ）. Conclusion miR -21 plays an important role in the proliferation and invasion of human laryngeal carcinoma cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第1期198-201,共4页
Chinese Journal of Experimental Surgery
基金
江苏省省级重点研发专项基金（BE2015666）
镇江市社会发展基金（SH2011059、SH2012042、SH2014027、SH2014047、SH2014050）
作者简介
通信作者：钱炜，Email：13338817788@163．com
