Construction of recombinant pEGFP-C1-Smad4 and its expression in human gastric cancer SGC7901 cell lines

Objective In order to make basis for gene therapy of gastric cancer, pEGFP-C1-Smad4 expression vector was constructed and we observed its expression in human gastric cancer SGC7901 cell lines. Methods Recombinant pEGFP-C1-Smad4 expression vector was constructed by techniques of gene recombination, screening and identified by restriction enzyme digestion and PCR. Then the recombinant was transfected into human gastric cancer cell line SGC7901 by techniques of gene transfection and detected expression by fluoroscopy and Western blot. Results Identification of pEGFP-C1-Smad4 by enzyme digestion and PCR showed that the length, inserted location and direction of the target gene inserted into the recombinant was correct and the expression of EGFP-C1 in transfected human gastric cancer SGC7901 cell was observed. The protein expression of Smad4 was detectable by Western blot. Conclusion The eukaryotic expression plasmid pEGFP-C1-Smad4 has been successfully constructed and it can be expressed transiently in SGC7901 cells, which would play a foundation for the further studies on the role of Smad4 in gastric carcinogenesis.