中国耳鼻咽喉头颈外科2016，Vol.23Issue(9)：507-510,514,5.DOI:10.16066/j.1672-7002.2016.09.004
RNA干扰技术沉默上皮型钙黏蛋白表达对体外培养的喉癌Hep-2细胞增殖的影响
Influence of RNAi on the silencing expression of E-cadherin and the proliferation ability of Hep-2 trained in vitro
孙静 1田军 2陈琪 3吴桂卿4
作者信息
- 1. 青岛市市立医院耳鼻咽喉科，山东 青岛 266000
- 2. 潍坊市人民医院耳鼻咽喉科，山东 潍坊 261000
- 3. 枣庄市市立医院耳鼻咽喉科，山东 枣庄 277000
- 4. 赣南医学院第一附属医院耳鼻咽喉科，江西 赣南 341099
- 折叠
摘要
Abstract
OBJECTIVE To explore the effect of E-cadherin on the proliferation ability of Hep-2 by method of RNA interference technology to silence the expression of E-cadherin. METHODS The specific siRNA sequences and non-silencing siRNA were designed and synthesized. Hep-2 cells were transfected and then the down expression of E-cadherin gene in vitro cultured Hep-2 cells were got. The silencing effect of E-cadherin gene was explored by Fluorescence Quantitative Polymerase Chain Reaction and the proliferation of the transfected Hep-2 cells were detected in vitro by MTT assay. RESULTS 1.When transfected with the ratio of recombinant plasmid and the quality of liposome volume at 1:1, the transfection efficiency at the siRNA-3 group was the highest and can be up to 65%. 2.The results of Fluorescence Quantitative Polymerase Chain Reaction: recombinant plasmid pRNAT-U6.1/Neo-siRNA1, pRNAT-U6.1/Neo-siRNA2 and pRNAT-U6.1/Neo-siRNA3 can down regulate the expression of E-cadherin mRAN. Set blank control group as a baseline (set to 1), the changes of expression of E-cadherin relative to β-actin in siRNA-1group was 0.00092, siRNA-2 group was 0.00143, siRNA-3 group was 0.00045 and the negative control group was 3.44898. The difference was statistically significant (P<0.05). 3. MTT: The growth rate of Hep-2 cells treated by specific siRNA was faster than that of the control group, and the difference was statistically significant (P<0.05). CONCLUSION Effectively inhibition the expression of E-cadherin's mRAN can enhance the proliferation of Hep-2 cells.关键词
RNA干扰/细胞增殖/喉肿瘤/钙粘蛋白Key words
RNA Interference/Cell Proliferation/Laryngeal Neoplasms/cadherin引用本文复制引用
孙静,田军,陈琪,吴桂卿..RNA干扰技术沉默上皮型钙黏蛋白表达对体外培养的喉癌Hep-2细胞增殖的影响[J].中国耳鼻咽喉头颈外科,2016,23(9):507-510,514,5.
