Vector construction and silencing effect of HER4 gene targeted small interfering RNA / 医学研究生学报

ABSTRACT

Objective RNA interference refers to post-transcriptional gene silencing caused by double strands RNA.To investigate the effect of EGFR receptor on esophageal carcinoma,the expression vector of HER4 gene targeted small interfering RNA was constructed to observe its silencing effect in human esophageal carcinoma cell line Eca-109,in order to find a promising method for the gene therapy of this disease.Methods Two complementary oligo DNA strands targeting HER4 gene were designed and synthesized according to the principles of designing siRNA.After annealing,oligo DNAs were inserted into SUPER.neo+gfp vector,then enzyme digestion analysis and DNA sequencing were applied.After transfecting it into human esophageal carcinoma cell line,we detected the level of expression of HER4 gene through real-time quantitative PCR and Western Blot.Results The enzyme digestion analysis and DNA sequencing show that HER4 gene targeted small interfering RNA and its expression vector were constructed successfully,and after transfection,the expression of HER4 gene in esophageal carcinoma cell line was suppressed greatly.Conclusion HER4 gene targeted small interfering RNA and its expression vector were constructed successfully,and could decrease the expression of HER4 gene in Eca-109 cell line,which laid the foundation for the following experiment.