Construction of pEGFP-C1-STAT3 Recombinant Eukaryotic Expression Vector and Its Effect on Expression of Snail in SGC7901 Cell Line

Objective To investigate the effect on the expression of zinc finger transcript factor Snail1(Snail) and excision repair cross complementation group 1 (ERCC1) after transfected with the signal transducer and activator of transcription factor(STAT3) in SGC7901 cell line. Methods We used recombinant DNA technology to construct the pEGFP-C1-STAT3 recombinant eukaryotic expression vector and transfected by liposome 2000 in SGC7901.The expression of EGFP was observed in transfection group by fluorescent microscopy.We detected the expression of STAT3,pSTAT3,Snail, ERCC1 and the apoptosis rate using Western blot and flow cytometry after cisplatin (DDP) treated. Results The recombinant plasmid was confirmed by double restriction enzyme digestion and the sequence was consensus with STAT3 gene sequencing.SGC7901 cells were transfected with recombinant plasmid pEGFP-C1-STAT3 and the product of recombinant plasmid was obtained.Western blot detection of the pSTAT3,Snail and ERCC1 expression showed significantly increased protein levels.Flow cytometry analysis revealed obviously decreasing of cell early apoptosis after the DDP treatment in pEGFP-C1-STAT3 transfection group. Conclusion We constructed the pEGFP-C1-STAT3 recombinant eukaryotic expression vector successfully and enhanced the expression of Snail and ERCC1 after transfection in SGC7901 cell line,which decreased its sensitivity to cisplatin.