Effects of Smad4 over-expression on proliferation of human gastric cancer cells and activation of NF-?B pathway / 中国肿瘤生物治疗杂志

ABSTRACT

Objective:To construct pEGFP-C1-Smad4 expression vector and to observe the influence of Smad4 over-expression on the proliferation of human gastric cancer SGC7901 cells and its relationship with nuclear factor kappa B (NF-?B). Methods:Recombinant expression vector pEGFP-C1-Smad4 was constructed and was used to transfect human gastric cancer SGC7901 cells. EGFP expression in transfected cells was detected by fluoroscopy. Smad4 and NF-?B expression in transfectant was examined by Western blotting. Effect of Smad4 over-expression on activation of NF-?B and proliferation of transfected SGC7901 cells were examined by electrophoretic mobility shift assay (EMSA) and MTT assay,respectively. Results:Expression of EGFP in transfected SGC7901 cells was observed under fluorescence microscope. Smad4 was over-expressed in transfected SGC7901 cells,accompanied by down-regulation of NF-?B p65 expression in the tranfectants. EMSA and MTT demonstrated that Smad4 over-expression significantly inhibited the activation of NF-?B and the proliferation of SGC7901 cells (P