Effects of COX-2 gene expression inhibited by shRNA on proliferation of gastric carcinoma SGC-7901 cells

Objective To investigate the inhibiting effects of shRNA(short hairpin RNA,shRNA) on COX-2 gene expression,the cell cycle and growth of gastric carcinoma SGC-7901 cells.Methods Specific shRNA plasmid to COX-2 were constructed,and then transfected into SGC-7901 cells by lipofectamine methods.Tests were divided into three groups: untransfected gastric carcinoma SGC-7901 cells group,negative control HK group and pshRNA-COX-2 group.Gastric carcinoma SGC-7901 cells were transfected with LipofectamineTM 2000.Expression of COX-2mRNA and protein were detected with reverse transcriptional polymerase chain reaction(RT-PCR) and Western blot respectively.Cell cycle analysis and cell growth chart were detected with flow cytometry and cell count respectively.Results Compared with negative control HK group,recombinant expression vector pshRNA-COX-2 resulted in the reduction of COX-2mRNA and protein expresion by 70.1% and 43.2% respectively;cells in G0-G1 phase increased from 61.5%to 70.2%,cells in S phase decreased from 27.3% to 21.7%,and the growth of SGC-7901 cells cells was slowed significantly.Conclusions Recombinant expression vector pshRNA-COX-2 can significantly inhibit the expression of COX-2 gene,result in the increase of cells in G0-G1 phase and decrease of cells in S phase,and suppress proliferation of gastric carcinoma SGC-7901 cells.