Influence of RNAi on the silencing expression of E-cadherin and the proliferation ability of Hep-2 trained in vitro
10.16066/j.1672-7002.2016.09.004
- VernacularTitle: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
- Author:
Jing SUN
;
Jun TIAN
;
Qi CHEN
;
Guiqing WU
- Publication Type:Journal Article
- Keywords:
RNA Interference;
Cell Proliferation;
Laryngeal Neoplasms;
cadherin
- From:
Chinese Archives of Otolaryngology-Head and Neck Surgery
2016;23(9):507-510,514
- CountryChina
- Language:Chinese
-
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.
