Abstract:Objective:To study the interfering effect of short hairpin RNA(shRNA) expressing plasmid vectors on STAT3 gene expression of human gastric carcinoma cells.
Methods :Specific shRNA plasmids to STAT3 were constructed, and then transfected into MKN-45 cells by lipofectamine methods. Semi-quantitative RT-PCR and Western blotting were used to detect the expression of STAT3 mRNA and protein respectively. Cells were divided into three groups: control group (A group), psiRNA-H1 group (negative group, B group) and psiRNA-H1/STAT3 group (C group).
Results:The synthesized strands of oligonucleotide contained correct and complete sequence of the shRNA, and the STAT3 specific shRNA was inserted into eukaryotic expression．Compared with the A and B group cells, semi-quantitative RT-PCR and Western blotting showed the expression of STAT3 mRNA and protein was down-regulated in the C group (P<0.05).
Conclusions:The recombinant plasmid psiRNA-H1/STAT3 shRNA has been constructed successfully, and it can specificly inhibit not only the expression of STAT3 mRNA, but also the protein expression in gastric carcinoma cells in vitro.It provides certain experimental basis for STAT3 gene target therapy.