Effect of shRNA interfering plasmid targeting FOXC1 gene on proliferation of gastric adenocarcinoma SGC-7901 cells

Objective To investigate the effect of shRNA interfering plasmid targeting FOXC1 gene on proliferation of gastric adenocarcinoma SGC-7901 cells.Methods The transcription level of FOXC1 mRNA and expression level of FOXC1 protein in SGC-7901 and gastric mucosal epithelium GES-1 cells were determined by RT-PCR and Western blot respectively.Three shRNA interfering plasmids targeting FOXC1 gene,i.e.pshRNA-FOXC1a,pshRNA-FOXC1b and pshRNA-FOXC1c,were constructed and transfected to SGC-7901 cells respectively,using the cells untransfected and those transfected with empty plasmid pGenesil as controls.The transfected SGC-7901 cells were determined for transcription level of FOXC1 mRNA and expression level of FOXC1 protein by RTPCR and Western blot respectively,for proliferation activity by MTT method,and for cell cycle by flow cytometry.Results Both the transcription level of FOXC1 mRNA and expression level of FOXC1 protein in SGC-7901 cells were significantly higher than those in GES-1 cells(P 0.05).The transcription level of FOXC1 mRNA and expression level of FOXC1 protein in SGC-7901 cells transf ected with pshRNA-FOXC1a,pshRNA-FOXC1b and pshRNA-FOXC1c were significantly lower than those transfected with pGenesil-1 and those untransfected(P 0.01),while the inhibition rate of proliferation was significantly higher than that transfected with pGensil-1(P 0.05).However,the percentages at G1 and G2 phases of SGC-7901 cells transfected with pshRNA-FOXC1a,pshRNAFOXC1b and pshRNA-FOXC1c were significantly higher,while that at S phase was significantly lower,than those untransfected(P 0.01).Conclusion The shRNA eukaryotic expression plasmid for FOXC1 gene was successfully constructed.FOXC1 was highly expressed in SGC-7901s,which arrested the cell cycle at G1 ~ G2 phase and inhibited the cell proliferation.